Fig 2: The effect of differentiation on MYOC levels and its localization in C2C12 cells. (A). Change in MYOC, MYH2, and MYOG expression levels during C2C12 cell differentiation. (B). Gray scanning of MYOC in Figure A. (C). Gray-level scanning of MYH2 in Figure A. (D). Gray scanning of MYOG in Figure A. (E). Immunofluorescence was used to measure changes in MYOC expression and the size and length of myotubes during the differentiation of C2C12 cells. (F). MYOC was labeled green with FITC, while the nucleus was stained blue with DAPI. MYOC was principally located on the cell membrane during the differentiation of C2C12 cells, as identified by laser confocal detection. (** p values < 0.01, * p values < 0.05) (n = 3).

Fig 3: MYOC exerts an effect on the differentiation of C2C12 cells by regulation of CAV1 that affects the TGF-ß pathway. (A). Change in MYOC, P-Smad2, and P-SMad4 expression after transfection of siRNA-sequence T3 for MYOC silencing. (B). Grayscale scan of MYOC in Figure A. (C). Grayscale scan of P-Smad2 in Figure A. (D). Grayscale scan of P-Smad4 in Figure A. (E). Western blotting of all indices in the co-transfection experiment control group and 3 experimental groups. (F). Grayscale scan of MYOC in Figure E. (G). Grayscale scan of CAV1 in Figure E. (H). Grayscale scan of MYH2 in Figure E. (I). Grayscale scan of MYOG in Figure E. (J). Grayscale scan of P-Smad2 in Figure E. (K). Grayscale scanning of P-Smad4 in Figure E. (L). Change in rate of fusion of myotubes after co-transfection, as measured by laser confocal microscopy. (M). Myotubule fusion rate in Figure L. (** p values < 0.01, * p values < 0.05) (n = 3).

Fig 4: Effects of CAV1 on C2C12 cell differentiation. (A). Change in CAV1, MYH2, and MYOG expression levels at different stages of C2C12 cell differentiation. (B). Grayscale scan of CAV1 in Figure A. (C). Grayscale scan of MYH2 in Figure A. (D). Grayscale scan of MYOG in Figure A. (E). Immunofluorescence was used to measure the changes in CAV1 expression levels and the morphology of myotubes at various stages of C2C12 differentiation. CAV1 was labeled green using FITC and the nucleus blue using DAPI. (F). Laser confocal microscopy demonstrating the location of CAV1 expression, stained red using RBFITC, and nuclei blue using DAPI. (G). Changes in CAV1, MYH2, and MYOG expression levels after siRNA transfection for 48 h and 72 h. (H). Grayscale scan of CAV1 in Figure G. (I). Grayscale scan of MYH2 in Figure G. (J). Grayscale scan of MYOG in Figure G. (K). Changes in the fusion rate of C2C12 cells in mice after CAV1 inhibition by immunofluorescence detection. (L). Myotubule fusion rate in Figure K. (M). Change in CAV1, P-Smad2, and P-Smad4 expression after siRNA fragment S1 transfection of CAV1. (N). Grayscale scan of CAV1 in Figure M. (O). Grayscale scan of P-Smad2 in Figure M. (P). Grayscale scan of P-Smad4 in Figure M. (** p values < 0.01, * p values < 0.05) (n = 3).