Supplementary Figure 4: Effects of manipulations of RhoA activity on TCP and rab11 interaction with RhoA signalling.
From: Transient junction anisotropies orient annular cell polarization in the Drosophila airway tubes
Confocal projections of the DT at 2nd instar larvae at 64–66 h AEL upon expression of a constitutive active form (a–c) or a dominant negative form (d–f) of RhoA. Flip-out clones using the Act5c>y>gal4 construct are marked by cytoplasmic GFP or CD8-GFP (green). DE-cad marks all junctions. RhoAV14 expression increases levels of pMLC (a) or phalloidin labelled F-actin cables (b) (arrows) while RhoAN19 expression decreases levels of pMLC (d) or phalloidin labelled F-actin cables (e) (arrows). In either situation, Rab11 is also mislocalised at the transverse junctions (c,f, arrows). We note that single cell clones of RhoAN19 do not change directions of annular-TCP (e compare with f), suggesting that neighbouring wild type cells can communicate the direction of annular-TCP. g, In rab11-RNAi expressing clones, preferential pMLC accumulation at longitudinal junctions is lost. Quantification is shown in h. (n = 29 for L and 20 for T junctions from 6 rab11-RNAi larva, NS, not significant by unpaired two-tailed Student’s t test). Error bar, s.e.m. See also Supplementary Table 3 for scatterplots. Scale bars: 20 μm. Images in a–c are representative of at least 4 larvae from ≥ 2 independent experiments. Images in d–f are representative of at least 3 larvae from 2 independent experiments. Data in g and h are aggregated from 3 independent experiments.
