1. Blockage of peroxisomal FAO induces differentiation on activated satellite cells.
Blockage of peroxisomal FAO induces differentiation on activated satellite cells. (A) Schematic of experimental design. Satellite cells were activated in vivo by injection of notexin into the tibialis anterior muscle of Tg:Pax7-nGFP mice and isolated at day 5 (D5). Cells were cultured for 48 h in presence or absence of treatment (same experimental design for drug treatment and siRNA transfection; n=4 per condition; T1, isolation; T2; analysis). Cells were pulsed with EdU for 4 h prior fixation. (B) Immunostaining for Pax7 (green), myogenin (red) and EdU (cyan) of untreated cells or cells treated with 5 µM Thioridazine or 5 µM Etomoxir for 48 h. Cells were pulsed with EdU 4 h prior to fixation. Hoechst staining shows nuclei. (C) Histogram representing percentage of cells positive for EdU and Myog (EdU+ and Myog+, respectively). (D) Expression levels of indicated genes were calculated and normalised to that of the reference gene ribosomal protein L13aRPL13a. Expression is shown as fold-change to non-treated control. Perox, peroxisomal genes; Mito, mitochondrial genes. (E) Immunostaining for EdU (green) and myogenin (red) of cells transfected with control siRNA or siRNA against catalase (siCAT), acyl-coenzyme oxidase 1 (siACOX1) and carnitine palmitoyltransferase 1b (siCPT1b). (F) Histogram representing percentage of EdU+ and Myog+ cells. (G) Expression levels of indicated genes relative to that of RPL13a and normalised to scramble control. Data are presented as mean (±s.e.m.). All P-values were calculated using Student’s t-test. *P<0.05, **P<0.01, ***P<0.001.
Francesca Pala et al. J Cell Sci 2018;131:jcs212977
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