Fig. 3

Attenuation of mTORC1 signaling in OPTN(E50K) hPSC-RGCs. A–F Western blot and the relative protein expression of pS6^Ser240/244, pAKT^Ser473 and pAMPK^Thr172 to its total protein, respectively, in hPSC-RGCs (n = 6 for each WT and E50K; t-test, pS6^Ser240/244 **p = 0.0057; pAKT^Ser473 ns = not significant, p = 0.854; pAMPK^Thr172 **p = 0.0027). G–M Immunostaining and quantification of pS6^Ser240/244 intensity revealed that a subset of RGCs with the OPTN(E50K) mutation reduced the expression of pS6^Ser240/244 (n = 3 biological replicates using WT n = 96 and E50K n = 96 technical replicates; t-test, ***p = 0.0009). Arrows indicate RGCs that have reduced expression of pS6^Ser240/244. N RNA-seq analyses demonstrated a significant reduction in some mTOR-associated genes. Scale bar: 25 μm. Data are all represented as mean values ± SEM