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. 2022 May 16;132(10):e156125. doi: 10.1172/JCI156125

Figure 3. Aerobic exercise modestly increased cytosolic content of MBNL2, but does not alter MBNL1 sequestration.

Figure 3

(A) Representative Western blot of p-AMPKThr172, t-AMPK, and PGC-1α in the vastus lateralis muscle. (B) Graphical summary of p-AMPKThr172, t-AMPK, and PGC-1α expression. (C) Representative Western blot of MBNL1, MBNL2, and CUGBP1. (D) Graphical summary of MBNL1, MBNL2, and CUGBP1 expression. (E) Representative Western blot of p-GSK3βSer9 and t-GSK3β. (F) Graphical summary of p-GSK3βSer9, t-GSK3β, and inhibition status (p-GSK3βSer9 relative to t-GSK3β) expression. A typical Ponceau stain displayed below demonstrates sample loading. (G) Representative Western blot of p-AMPKThr172, t-AMPK, MBNL1, MBNL2, and CUGBP1 in nuclear and cytosolic fractions from DM1-PRE and DM1-POST. Histone 3 and GAPDH proteins displayed below to indicate nuclear and cytosolic fraction purity. Approximate molecular weights (kDa) shown at right of blots in A, C, and E. (H–K) Graphical summary of p-AMPKThr172, t-AMPK, MBNL1, and MBNL2 in nuclear and cytosolic fractions. (L) Representative images of combined FISH probing for CUG repeats ([CUG]n) and IF staining of MBNL1 along with DAPI to mark myonuclei and merged image. Original magnification, ×60 with a ×10 digital imaging zoom, for final magnification of ×600. (M) Summary of the numbers of MBNL1/(CUG)n-positive myonuclei as an indicator of MBNL1 sequestration. (N) Average number of foci within MBNL1/(CUG)n-positive myonuclei. Data are expressed as box and whisker plots with plus signs representing the mean (B–F, M, and N) or bar graphs as mean ± SEM (H–K). n = 9–11. *P < 0.05 versus CON, 1-way ANOVA followed by Bonferroni’s correction; #P < 0.05 versus DM1-PRE, 2-tailed paired t test corrected for multiple comparisons.