Skip to main content
. 2019 Apr 15;129(5):2088–2093. doi: 10.1172/JCI122560

Figure 3. LRS inhibits myogenic differentiation through the Rag-mTORC1 pathway.

Figure 3

(A) C2C12 cells were transduced with lentiviruses expressing shLRS or shScramble, and puromycin-selected for 2 days. Cells were then transfected with either empty vector (EV) or HA-GST-RagB-Q99L and HA-GST-RagD-S77L for 24 hours, differentiated for 72 hours, and subjected to measurement of differentiation index. Data were normalized to control (shScramble and empty vector) (n = 3). (B) LRS was transfected in cells where RagA and RagB were knocked down and subjected to measurement of differentiation index as in A (n = 3). (C) LRS was transfected in cells where Raptor was knocked down and subjected to measurement of differentiation index as in A (n = 4). (D) Cells were transduced with lentiviruses expressing shLRS or shScramble, and puromycin-selected for 2 days. Upon confluence, cells were lysed and subjected to Western blotting analysis (n = 2). (E, F) Cells were transfected with myr-Akt and/or LRS. Confluent cells were either induced to differentiate for 72 hours, followed by immunofluorescence staining with anti-MHC (green) and DAPI (blue) (E, n = 3), or lysed for Western blotting analysis (F, n = 3). Scale bar: 50 μm. The data denoted by #, §, ‡ are significantly different from each other by 2-way ANOVA (P < 0.005). All error bars represent SEM.