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. 2018 Oct 11;10(11):e8799. doi: 10.15252/emmm.201708799

Figure 4. Rapamycin induces autophagy in Cox15 sm/sm muscles.

Figure 4

  • A
    Representative Western blot of autophagy markers at steady state in skeletal muscle. GAPDH was used as a loading control.
  • B
    Densitometric analysis of immunoblots similar to that shown in (A) (n = 4/group). Black: untreated WT; gray: rapamycin‐treated WT; red: untreated Cox15 sm/sm; blue: rapamycin‐treated Cox15 sm/sm. Error bars represent SEM. The asterisks represent the significance levels calculated by one‐way ANOVA with Tukey's post hoc multiple comparison test: **P = 0.0068 (LC3‐II WT versus Cox15 sm/sm), **P = 0.0063 (P62 WT versus Cox15 sm/sm), *P = 0.025 (P62 Cox15 sm/sm versus Cox15 sm/sm+rapamycin), **P = 0.0098 (S6 WT versus Cox15 sm/sm), *P = 0.034 (P‐S6 WT versus WT+rapamycin), ***P = 0.0002 (P‐S6 Cox15 sm/sm versus Cox15 sm/sm+rapamycin), *P = 0.014 (P‐S6/S6 WT versus WT+rapamycin), ***P = 0.0007 (P‐S6/S6 Cox15 sm/sm versus Cox15 sm/sm+rapamycin).
  • C
    Analysis of the autophagic flux (n = 3/group). Upper panel: representative Western blot of Cox15 sm/sm and WT muscles treated with colchicine, rapamycin, or rapamycin plus colchicine. Note that LC3‐II was increased in Cox15 sm/sm versus WT samples. Colchicine did not increase LC3‐II levels, suggesting a block in the autophagic flux; rapamycin plus colchicine‐treated Cox15 sm/sm mice showed higher levels of LC3‐II, suggesting that rapamycin increased the autophagic flux in Cox15 sm/sm muscles. Black: WT; dark gray: colchicine‐treated WT; middle gray: rapamycin‐treated WT; light gray: colchicine plus rapamycin‐treated mice; red: untreated Cox15 sm/sm; orange: colchicine‐treated; blue: rapamycin‐treated Cox15 sm/sm; purple: colchicine plus rapamycin‐treated Cox15 sm/sm. Error bars represent SEM. The asterisks represent the significance levels calculated by one‐way ANOVA with Tukey's post hoc multiple comparison test: **P = 0.0049 (WT versus WT+colchicine), **P = 0.0046 (WT+rapamycin versus WT+rapamycin+colchicine), *P = 0.0225 (WT versus Cox15 sm/sm), **P = 0.0057 (Cox15 sm/sm colchicine versus Cox15 sm/sm+rapamycin+colchicine), ***P = 0.0003 (Cox15 sm/sm rapamycin versus Cox15 sm/sm+rapamycin+colchicine).