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. 2018 Jun 4;217(6):2005–2018. doi: 10.1083/jcb.201708137

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© 2018 Wang et al.

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Figure 4. — Myonuclear DNA content does not change in myofibers with disrupted microtubules. Representative single confocal stacks of third-instar larvae muscle 7 in control (A–C) or after muscle-specific, temporal expression of the MT-severing protein Spastin induced for 2 d during second to third-instar larval growth stages (D–F). Muscles were labeled with anti–α-Tubulin (green; A, C, D, and F), Hoechst (blue), and Phalloidin (red; B, C, E, and F). C and F are merged images. Bar, 10 µm. Note the deletion of α-Tubulin labeling in the Spastin-expressing muscles and its normal expression in the trachea (Tr., arrow; D). Ortho view shown in C′ and F′ corresponds to the white lines in C and in F. (G) Quantitative analysis of the mean myonuclear area in control (Mef2-GAL4;TubGAL80/yw) versus MT-depleted muscle 7 during second- to third-instar stage (Mef-GAL4;TubGAL80>UAS-Spastin) indicates higher nuclear area in the latter group (t test: ****, P = 6 × 10⁻¹²). (H) Quantitative analysis of the mean myonuclear Hoechst integrated density in control (Mef2-GAL4;TubGAL80/yw) versus MT-deleted muscle 7 during second- to third-instar stage (Mef-GAL4;TubGAL80>UAS-Spastin) indicates no significant difference between the two groups (t test: P = 0.13). Images were taken from five different larvae (control, n = 17; Spastin RNAi. n = 18). Whiskers in G and H extend to data points less than 1.5 interquartile ranges from the first and third quartiles.