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. 2021 Apr 15;131(8):e135963. doi: 10.1172/JCI135963

Figure 10. Disruption of caveolae is sufficient to normalize insulin signaling with LPCAT3 knockdown.

Figure 10

C2C12 myoblasts were infected with lentivirus expressing shRNA for scrambled (SC) or LPCAT3-knockdown (KD) sequences and were differentiated into myotubes. (A–D) Myotubes were incubated with or without 10 mM of methyl-β-cyclodextrin (MβCD) for 1 hour. (A and B) MβCD successfully depletes cav3 (P < 0.001, main effect of LPCAT3 knockdown) but not flotillin-1 (P = 0.003, main effect of LPCAT3 knockdown; P = 0.01, main effect of MβCD) (n = 9). (C and D) Cells were incubated in the presence (0.6 nM) or absence of insulin and were blotted for total or Ser473 phosphorylation of Akt (n = 3 basal, n = 6 insulin). (E and F) C2C12 myotubes were incubated in the absence or presence of either PDGF (2.5 ng/mL), EGF (100 ng/mL), or insulin (12 nM) and were blotted for total or Ser473 phosphorylation of Akt (basal and insulin, n = 8; PDGF and EGF, n = 10). Two-way ANOVA with Šidák’s multiple-comparison test was performed. All data are represented as mean ± SEM. *P ≤ 0.05.