Fig. 4
Methylene blue and intermittent fasting preserve HDAC4mKO skeletal muscle structure following denervation. a Hematoxylin and eosin staining of HDAC4mKO contralateral and denervated muscles, 4 weeks after denervation, without (−) or after MB or IF treatment. Scale bar = 50 μm. b Number of myofibers in HDAC4mKO contralateral and denervated muscles, 4 weeks after denervation, without (−) or after MB or IF treatment. n = 3. Number of HDAC4mKO myofibers with 2500–3000 or 1000–1500 μm2 cross-sectional area, without (−) or after MB or IF treatment, 4 weeks after denervation. Data are shown as mean ± SEM; n = 4; two-way ANOVA revealed an interaction (F = 6.72; df 1; p = 0.02 for MB; F = 8.2; df 1; p = 0.01 for IF) (F = 7.27; df 1; p = 0.026 for MB; F = 10.2; df 1; p = 0.01 for IF); **p < 0.01 by Tukey’s HSD test. c HDAC4mKO muscle sections labeled with laminin (green) and IgG (red), 4 weeks after denervation, without (−) or after MB or IF treatments, with relative quantification. Scale bar = 50 μm. n = 3; two-way ANOVA showed an interaction (F = 10.82; df 1; p = 0.0094 for MB; F = 11.43; df 1; p = 0.0081 for IF); *p < 0.05 by Tukey’s HSD test. d Immunostaining for CD68 (green) and laminin (red) of HDAC4mKO muscles 4 weeks after denervation, without (−) or after MB or IF treatment, with relative quantification. Scale bar = 25 μm. n = 3; two-way ANOVA showed an interaction (F = 6.57; df 1; p = 0.03 for MB; F = 6.06; df 1; p = 0.036 for IF); *p < 0.05 by Tukey’s HSD test. e Masson’s trichrome staining of HDAC4mKO contralateral and denervated muscles, 4 weeks after denervation, without (−) or after MB or IF treatment, with relative quantification. Scale bar = 50 μm. n = 3; two-way ANOVA showed an interaction (F = 53.95; df 1; p = 0.001 for MB and F = 59.36; df 1; p = 0.0001 for IF); *p < 0.05 by Tukey’s HSD test.