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. 2021 Sep 10;7(37):eabi8787. doi: 10.1126/sciadv.abi8787

Fig. 4. Inhibition of TGFβ signaling in DMD-R3381X neuromuscular circuits rescues their myofiber contractility defect.

Fig. 4.

(A) Relative expression levels of TGFB1 transcripts in 2D myogenic cultures of DMD-R3381X, DMD-R3381X + SB-431542, and CORR-R3381X at 0, 24, and 120 hours of secondary differentiation. n = 6. Values are means ± SEM. One-way ANOVA and Tukey’s multiple comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. (B) Quantification of titin-positive cells in 2D myogenic cultures of DMD-R3381X, DMD-R3381X + SB-431542, and CORR-R3381X at 120 hours of secondary differentiation (representative images in fig. S4C). n = 4. Values are means ± SD. One-way ANOVA and Tukey’s multiple comparisons test, **P < 0.01. (C) PIV analysis in central compartments containing DMD-R3381X neuromuscular circuits not treated and treated with SB-431542 on day 5. Scale bars, 250 μm. (D) Quantification of mean velocity of DMD-R3381X myofibers not treated and treated with SB-431542 in response to optogenetic stimulation. n = 12. Values are means ± SEM. Two-way ANOVA and Sidak’s multiple comparisons test, **P < 0.01, ***P < 0.001, and ****P < 0.0001. (E) PIV analysis in central compartments containing CORR-R3381X neuromuscular circuits not treated and treated with SB-431542 on day 5. Scale bars, 250 μm. (F) Quantification of mean velocity of CORR-R3381X myofibers not treated and treated with SB-431542 in response to optogenetic stimulation. n = 12. Values are means ± SEM. Two-way ANOVA and Sidak’s multiple comparisons test, no significant difference. (C and E) Representative images show before, during, and after optogenetic stimulation. Green arrows represent velocity vectors. (D and F) Blue shading indicates the time during optogenetic stimulation.