Fig. 1: Spatiotemporal regulation of ADAMTSL2 during muscle regeneration.
From: Myoblast-derived ADAMTS-like 2 promotes skeletal muscle regeneration after injury
a Experimental design for BaCl2 injury in wild type (WT) mice. b, c Representative images of cross-sections of uninjured control (Ctrl) and injured tibialis anterior (TA) muscles at 7 (b) and 14 days post injury (dpi) (c). Sections were stained for ADAMTSL2 (red) and laminin (green). Nuclei were stained with DAPI (blue). d Quantification of ADAMTSL2 mean fluorescence intensity at 7 and 14 dpi (n = 6 mice, 1-2 fields-of-view). e Quantification of myofiber cross-sectional area (CSA) at 7 and 14 dpi (n = 6 mice, number of myofibers indicated). f Western blot detection of ADAMTSL2 and GAPDH in Ctrl and injured TA muscle extracts at 7 and 14 dpi. g Quantification of fluorescence intensities of ADAMTSL2 bands normalized to GAPDH (n = 3-4 mice). h qRT-PCR quantification of Adamtsl2 mRNA at 4 and 14 dpi normalized to Gapdh (n = 3 mice). i Representative images of TA muscles cross-sections of 8-week-old D2 Ctrl and D2.mdx mice. Sections were stained for ADAMTSL2 (red) and laminin (green). Nuclei were stained with DAPI (blue). j Quantification of ADAMTSL2 mean fluorescence intensities from D2 and D2.mdx TA muscles (n = 3 mice). k qRT-PCR quantification of Adamtsl2 mRNA normalized to Gapdh (n = 4 mice). l Analysis of muscle regeneration after cardiotoxin injury. The percentage of positive cells per gene is shown. Total number of analyzed cells/time point: 0 = 5670, 0.5 dpi = 3510, 2 dpi = 8395, 3.5 dpi = 11251, 5 dpi = 9491, 10 dpi = 6214, 21 dpi = 8662. m, n Expression levels of Adamtsl2 (m) and Pax7 (n) after cardiotoxin injury. Data points represent normalized and scaled mRNA levels from individual cells. Cells with an expression value of 0 are not shown. Bars in d, g, h, j, k represent mean values and whiskers the standard deviation. Boxes in m, n represent 25th – 50th percentile range, whiskers in e, m, n the standard deviation, and horizontal lines the mean value. Statistical significance in d, e, g, h, j, k was calculated using a Student’s t test and in m, n using a one-way ANOVA with posthoc Tukey test. Scale bars in b, c, i = 50 μm. Graphs were generated using CorelDRAW and OriginPro.
