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. 2021 Nov 2;23(1):e53231. doi: 10.15252/embr.202153231

Figure 5. Daw signaling mediates the effects of Btz on NMJ size.

Figure 5

  • A
    Log2 fold changes in daw, ltl, and frac mRNA in btz2 /Df(3R)BSC497 larval carcasses compared to btz2 /+, measured by RNA‐Seq (blue) or qRT–PCR (green). **q < 0.01; *q < 0.05 by unpaired t‐test corrected for false discovery rate with the Benjamini–Hochberg procedure (RNA‐Seq) or **P < 0.01; *P < 0.05 by unpaired t‐test (qRT–PCR). n = 3 for each sample. Error bars show mean ± SD.
  • B–K
    Confocal images of the NMJ on larval muscles 6 and 7 in segment A3, stained with anti‐HRP (magenta) and anti‐P‐Mad (green in B, C) or anti‐Dlg (green in D‐K). (B) btz2 /+ control; (C) btz2 /Df(3R)BSC497. Insets show enlargements of the boxed regions stained with anti‐P‐Mad. Synaptic P‐Mad is lost in btz mutants. (D) UAS‐daw RNAi/+ control; (E) BG380‐GAL4/+; UAS‐daw RNAi/+; (F) mhc‐GAL4/UAS‐daw RNAi; (G) UAS‐daw RNAi/+; repo‐GAL4/+; (H) UAS‐daw/+; btz2 /Df(3R)BSC497; (I) tub‐GAL4/UAS‐daw; btz2 /Df(3R)BSC497; (J) tub‐GAL4/UAS‐btz; btz2 /Df(3R)BSC497; (K) tub‐GAL4/UAS‐daw. Scale bars, 30 μm (shown in B for B‐C and in D for D‐K). Scale bar for insets in (B, C), 10 μm (shown in B).
  • L
    Quantifications of the number of boutons normalized to muscle surface area (×1,000) in the indicated genotypes. ****P < 0.0001; ns, not significant by unpaired t‐test or Mann–Whitney test. n = 19 (UAS‐daw RNAi/+), n = 9 (BG380>daw RNAi; btz/Df; tub>daw), n = 23 (mhc>daw RNAi), n = 10 (repo>daw RNAi; tub>daw), n = 12 (btz/Df; UAS‐daw), or n = 14 (btz/Df; tub>btz). Error bars show mean ± SEM. Knocking down daw in motor neurons, muscles, or glia reduces NMJ size. Expressing UAS‐daw with tub‐GAL4 rescues NMJ size in btz mutants as efficiently as expressing UAS‐btz, but has no significant effect on NMJ size in a wild‐type background.

Source data are available online for this figure.