Fig. 5: Diphenhydramine in dimenhydrinate promotes muscle differentiation by inducing autophagy. | Experimental & Molecular Medicine

Fig. 5: Diphenhydramine in dimenhydrinate promotes muscle differentiation by inducing autophagy.

From: Transcriptome-based deep learning analysis identifies drug candidates targeting protein synthesis and autophagy for the treatment of muscle wasting disorder

Fig. 5

a Immunoblotting for MyoD, myogenin, and MHC, which are muscle differentiation markers, during the 7-day differentiation of C2C12 cells. b Representative microscopy images of C2C12 cell differentiation following treatment with varying concentrations of DH at different time points. c Immunoblotting for MyoD, myogenin, ATG5 and LC3. Chloroquine (CQ; 30 μM, 24 h) was used as an autophagy inhibitor. d Immunoblotting for mTOR-independent LC3 via dimenhydrinate after 1 day of differentiation with or without CQ or DH. e–g The treatment concentrations for 8-CT, DPH, and DH were uniformly set at 10 μM. e Confirmation of LC3 induction by target chemicals with or without amino acids. Torin-1 (5 μM, 6 h) was utilized as an mTOR pathway inhibitor. f Immunofluorescence staining image of LC3 after individual treatment with the three chemicals. Scale bars = 50 μm. g Relative intensity of LC3 in C2C12 cells after treatment with the three chemicals. The mean fluorescence intensity was measured using ImageJ. ns not significant, *p < 0.05, **p < 0.01, ***p < 0.005. Error bars: standard deviation (SD). DH dimenhydrinate, 8-CT 8-chlorotheophylline, DPH diphenhydramine.

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