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Supplementary TablesTable S1 Basic demographics and the origin of Hamstring tendon tissue samples for human tenocytes (hTeno) cultures and other experiments.Donor numberAge (Years)GenderExperiments131MaleImmunofluorescence staining for INSR-β, GLUT1 and GLUT4.2-NBDG glucose uptake assay.224Male333Male423MaleTotal collagen assay.COL-I ELISA.521Male626MaleTable S2 Normality test for the data analysis for 2-NBDG uptake assay, total collagen expression levels and COL-I expression levels.ExperimentDataNormality test performedp-value for normality testDownstream data analysis performed2-NBGD uptake assayNormalized datasetKolmogorov-Smirnov testp<0.001Mann-Whitney?testFold change datasetKolmogorov-Smirnov testp<0.001Kruskal–Wallis & Mann-Whitney U testTotal collagen assayNormalized datasetKolmogorov-Smirnov testp<0.001Mann-Whitney?testFold change datasetKolmogorov-Smirnov test0.160One-Way ANOVA & Tukey Post-Hoc testType I Collagen ELISA assayNormalized datasetKolmogorov-Smirnov testp<0.001Mann-Whitney?testFold change datasetKolmogorov-Smirnov test0.200One-Way ANOVA & Tukey Post-Hoc testTable S3 The p-values for 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose (2-NBDG) uptake in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM). The p-values for Mann-Whitney test for relative 2-NBDG uptake levels between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with or without insulin supplement compared to the basal group (untreated; without both TNF-?α and insulin). The significant p-values were shown in italic.Groupsp-value vs baseline control group (untreated)Basal group + insulin?0.0020.008 ?M TNF-α?0.0020.08 ?M TNF-α?0.0020.8 ?M TNF-α?0.0028 ?M TNF-α?0.0020.008 ?M TNF-α + insulin?0.0020.08 ?M TNF-α + insulin?0.0020.8 ?M TNF-α + insulin?0.0028 ?M TNF-α + insulin?0.002The p-values for Mann-Whitney test for the pairwise comparisons of the relative 2NBDG uptake levels of hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with insulin supplement compared to their corresponding groups without insulin supplement. The significant p-values were shown in italic.Basal group + insulin0.008 ?M TNF-α + Insulin0.08 ?M TNF-α + Insulin0.8 ?M TNF-α + Insulin8 ?M TNF-α + InsulinBasal group §0.0020.008 ?M TNF-α0.1800.08 ?M TNF-α0.1320.8 ?M TNF-α0.8188 ?M TNF-α0.818The p-values for Kruskal Wallis and Mann Whitney U for the fold change of insulin-mediated 2-NBDG upateke in hTeno treated with different concentrations of TNFα (0.008, 0.08, 0.8 and 8 μM) versus the non-TNF-α treated basal group. The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_glucose_uptakeChi-Square19.832df4Asymp. Sig..001a. Kruskal Wallis Testb. Grouping Variable: groupGroupBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group?0.004?0.004?0.004?0.0040.008 ?M TNF-α?0.0040.3370.0550.08 ?M TNF-α0.337‖0.0250.8 ?M TNF-α0.8738 ?M TNF-αTable S4 The p-values for total collagen expression levels in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM). The p-values for Mann-Whitney test for the relative total collagen expression levels between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with or without insulin supplement compared to the basal group (untreated; without both TNF-?α and insulin). The significant p-values were shown in italic.Groupsp-value vs basal groupBasal group + insulin?0.0010.008 ?M TNF-α?0.0020.08 ?M TNF-α ?0.0020.8 ?M TNF-α?0.0028 ?M TNF-α?0.0020.008 ?M TNF-α + insulin*0.0400.08 ?M TNF-α + insulin?0.0020.8 ?M TNF-α + insulin?0.0028 ?M TNF-α + insulin?0.002The p-values for Mann-Whitney test for the pairwise comparison of the relative total collagen expression levels of hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with insulin supplement compared to to their corresponding groups without insulin supplement. The significant p-values were shown in italic.Basal group + insulin0.008 ?M TNF-α + Insulin0.08 ?M TNF-α + Insulin0.8 ?M TNF-α + Insulin8 ?M TNF-α + InsulinBasal group§0.0020.008 ?M TNF-α§0.0040.08 ?M TNF-α0.1500.8 ?M TNF-α0.3378 ?M TNF-α§0.004The p-values for ANOVA and Tukey Post-Hoc test for the fold change of insulin-mediated total collagen expression levels in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) versus the non-TNF-α treated basal group. The significant p-values were shown in italic.ANOVAinsulin_mediated_TCE Sum of SquaresdfMean SquareFSig.Between Groups5.37441.34459.250.000Within Groups.56725.023Total5.94129p-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group?p<0.001‖0.011?p<0.001?p<0.0010.008 ?M TNF-α‖0.0140.420?p<0.0010.08 ?M TNF-α0.436?p<0.0010.8 ?M TNF-α?p<0.0018 ?M TNF-αTable S5 The p-values for type I collagen (COL-I) expression levels in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM). The p-value for Mann-Whitney test for the relative COL-I expression levels between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with or without insulin supplement compared to the basal group (untreated; without both TNF-?α and insulin). The significant p-values were shown in italic.Groupsp-value vs basal groupBasal group + insulin?0.0020.008 ?M TNF-α?0.0020.08 ?M TNF-α ?0.0020.8 ?M TNF-α?0.0028 ?M TNF-α0.3050.008 ?M TNF-α + insulin?0.0020.08 ?M TNF-α + insulin?0.0020.8 ?M TNF-α + insulin*0.0408 ?M TNF-α + insulin?0.002The p-values for Mann-Whitney test for the pairwise comparisons of the relative COLI expression levels of hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) with insulin supplement compared to their corresponding groups without insulin supplement. The significant p-values were shown in italic.Basal group + insulin0.008 ?M TNF-α + Insulin0.08 ?M TNF-α + Insulin0.8 ?M TNF-α + Insulin8 ?M TNF-α + InsulinBasal group §0.0020.008 ?M TNF-α0.7490.08 ?M TNF-α§0.0040.8 ?M TNF-α§0.0068 ?M TNF-α§0.004The p-values for ANOVA and Tukey Post-Hoc test for the fold change of insulin-mediated COL-I expression levels in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) versus the non-TNF-α treated basal group. The significant p-values were shown in italic.ANOVAinslin_mediated_COLI Sum of SquaresdfMean SquareFSig.Between Groups.5184.12911.305.000Within Groups.28625.011Total.80429p-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group?p < 0.0011.0000.0670.4220.008 ?M TNF-α?p < 0.0010.060?0.0050.08 ?M TNF-α‖0.0490.3410.8 ?M TNF-α0.8338 ?M TNF-αTable S6 The p-values for insulin-mediated normalized tenogenic-related markers (SCX and MKX) gene expressions in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM). The p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized SCX gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_SCXChi-Square22.836df4Asymp. Sig..000a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.3940.5220.0040.08 ?M TNF-α0.7490.0040.8 ?M TNF-α0.0048 ?M TNF-αThe p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized MKX gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_MKXChi-Square14.084df4Asymp. Sig..007a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.8730.8730.7490.08 ?M TNF-α0.8731.0000.8 ?M TNF-α0.8738 ?M TNF-αTable S7 The p-values for insulin-mediated normalized ECM-related markers (COL1A1, COL3A1, MMP-9, MMP-13) gene expressions in hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM). The p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized COL1A1 gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_COL1A1Chi-Square19.021df4Asymp. Sig..001a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.3370.1090.1090.08 ?M TNF-α0.0100.1090.8 ?M TNF-α0.3378 ?M TNF-αThe p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized COL3A1 gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,b insulin_mediated_COL3A1Chi-Square14.370df4Asymp. Sig..006a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.0550.2620.5220.08 ?M TNF-α0.3370.3370.8 ?M TNF-α0.0558 ?M TNF-αThe p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized MMP-9 gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_MMP9Chi-Square17.586df4Asymp. Sig..001a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.0780.0550.0550.08 ?M TNF-α1.0000.8730.8 ?M TNF-α1.0008 ?M TNF-αThe p-value for Kruskal-Wallis and Mann-Whitney tests for the insulin-mediated normalized MMP-13 gene expressions between hTeno treated with different concentrations of TNF-α (0.008, 0.08, 0.8 and 8 μM) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,binsulin_mediated_MMP13Chi-Square21.939df4Asymp. Sig..000a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α0.08 ?M TNF-α0.8 ?M TNF-α8 ?M TNF-αBasal group*0.002*0.002*0.002*0.0020.008 ?M TNF-α0.0250.0040.0040.08 ?M TNF-α0.3370.7490.8 ?M TNF-α0.2628 ?M TNF-αTable S8 The p-values for percentage of live and apoptotic cells in 0.008 μM treated-hTeno treated at different timepoints.The p-value for Kruskal-Wallis and Mann-Whitney tests for the percentage of live cells in 0.008 μM treated-hTeno treated at different timepoints (24h, 48h and 72h) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,bpercentage_live_cellsChi-Square19.057df3Asymp. Sig..000a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α (24h)0.008 ?M TNF-α (48h)0.008 ?M TNF-α (72h)Basal group*0.002*0.002*0.0020.008 ?M TNF-α (24h)0.1090.0040.008 ?M TNF-α (48h)0.0250.008 ?M TNF-α (72h)The p-value for Kruskal-Wallis and Mann-Whitney tests for the percentage of apoptotic cells in 0.008 μM treated-hTeno treated at different timepoints (24h, 48h and 72h) compared to the basal group (untreated). The significant p-values were shown in italic.Test Statisticsa,bapoptotic_cellsChi-Square19.395df3Asymp. Sig..000a. Kruskal Wallis Testb. Grouping Variable: groupsp-valueBasal group0.008 ?M TNF-α (24h)0.008 ?M TNF-α (48h)0.008 ?M TNF-α (72h)Basal group*0.002*0.002*0.0020.008 ?M TNF-α (24h)0.0540.0040.008 ?M TNF-α (48h)0.0250.008 ?M TNF-α (72h) ................
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