Fig. 4

Rapamycin and rapamycin plus metformin inhibited mTORC1 but had no effect on mTORC2 or autophagy. IHC was performed on the medial tibia for P-RPS6 (A; n = 7 per group) and quantified as percent positive cells (B) and mean integrated intensity (C). Red arrowheads indicate cells staining positive for P-RPS6. Western blot was performed on glenohumeral cartilage (D) for P-RPS6 (E), P-Akt (F), P-AMPK (G), LC3B (H), ADAMTS5 (I), and MMP13 (J). Data analyzed using a one-way ANOVA to detect differences between groups for all variables except P-RPS6 (cells+), P-Akt, P-AMPK, LC3B, ADAMTS5, and MMP13, for which differences were determined by the non-parametric Kruskal-Wallis test with Dunn’s multiple comparison test. n = 8 per group for Rap and n = 7 per group for Con and Rap+Met. Images are outlined in black to show that, while each band is from the same blot, bands were selected for presentation to best represent the mean change. *P < 0.05 vs Con, **P < 0.01 vs Con