Fig. 1: Cultured fibers express genes of atrophy and autophagy, but not of apoptosis.
From: Active acetylcholine receptors prevent the atrophy of skeletal muscles and favor reinnervation
Unilateral sciatic nerve transections were performed in Cx43fl/flCx45fl/fl mice (control mice); at day 7 post denervation the innervated (Inne) and denervated (Den) flexor digitorum brevis muscles were collected to determine the relative levels of specific mRNAs by qPCR (In vivo). In parallel experiments, myofibers from control mice were cultured for 0, 48, and 72 h under control conditions or treated with 200 nM carbachol (Cbc) or 100 nM Paclitaxel to determine the relative levels of specific mRNAs by qPCR and proteins by immunofluorescence (In vitro). a mRNA relative levels of Fbxo32 (atrogin-1), Trim63 (MuRF1), and Bnip3 normalized to actin β expression. b Relative levels of atrogin-1 (Green) and µ-calpain (Red) evaluated by immunofluorescence using confocal microscopy. Calibration bar: 50 µm. c mRNA relative levels of Bak1 (pro-apoptotic gene) and Bcl2 (anti-apoptotic gene) normalized to actin β expression. d Ratio of Bak1/Bcl2. e Relative levels of anexin V or caspase 3 (Green) evaluated by immunofluorescence (DAPI to stain nuclei in blue). Calibration bar: 50 µm. Bar error is the mean ± SEM. N = 4 independent experiments.
