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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: Muscle regulates mTOR dependent axonal local translation in motor neurons via CTRP3 secretion: implications for a neuromuscular disorder, spinal muscular atrophy

Fig. 5

CTRP3 activates the PI3K/mTOR and MAPK/ERK pathway in motor neurons. a-e Representative Western blots and quantification of AKT and ERK activity in NSC-34 cells treated with 5 μg/ml mCTRP3 for 5, 15, 30 or 60 min. Activity of PI3K and MAPK/ERK pathways were measured using anti-p-ERK (Thr202/Tyr204) and anti-p-AKT (S473) antibodies. (n = 3) f-j WT and k-o SMA motor neurons treated with 5 μg/ml mCTRP3 for 5, 15, 30 or 60 min. Activity of PI3K/mTOR and MAPK/ERK pathways were measured using anti-p-S6K (Thr389) and anti-p-ERK (Thr202/Tyr204) antibodies. ACTB was used as a loading control. g-j, l-o Dot plot bar graphs summarize the repeated experiments. Each dot represents an independent experiment and bar graphs depict the mean ± s.d. (WT: n = 11 for p-ERK and n = 9 for p-S6K, SMA: n = 6 for p-ERK n = 8 for p-S6K) Kruskal Wallis and Uncorrected Dunn’s test was used to determine statistical significance; ** p < 0.01 and * p < 0.05. p Differentiated NSC-34 cells were treated with 100 nM of water-soluble mTOR inhibitor WYE-687 dihydrochloride and/or 5 μg/ml human CTRP3 (hCTRP3) for 2 h. q Representative Western blots of NSC-34 cells with anti-p-S6 (Ser235/236), anti-S6 and anti-SMN antibodies show that elevated SMN levels by CTRP3 treatment were mTOR dependent. ACTB was used as a loading control. r Dot plot bar graphs summarize the repeated experiments. Each dot represents an independent experiment and bar graphs depict the mean ± s.d. Kruskal Wallis and Uncorrected Dunn’s test was used to determine statistical significance; ** p < 0.01 and * p < 0.05

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