Supplementary Figure 5: Transverse aPKC localization upon knock down of chic, RhoA or Rab11. Uif involvement in annular-TCP.

Confocal projections of the DT of 2nd instar larvae (63 h AEL) showing the transverse aPKC localization (arrows) in the wild type (a), chic-RNAi (b), RhoA^N19 (c) or rab11-RNAi. Flipped out clones of the Act5c>y>gal4 construct are marked with CD8-GFP (bottom) in c and d. e–g, Quantification of relative intensity of aPKC along longitudinal (L) or transverse junctions (T) in chic-RNAi (e), RhoA^N19 (f) or rab11-RNAi (g) (n = 43 for L and 37 for T junctions from 6 chic-RNAi larvae, ^∗∗∗∗P < 0.0001, n = 31 for L and 37 for T junctions from 6 RhoA N19 larvae, ^∗∗∗∗P < 0.0001, n = 26 for L and 17 for T junctions from 4 rab11-RNAi larvae, ^∗∗∗P = 0.0009 p-values were calculated using the unpaired two-tailed Students t-test). Error bar: s.e.m. Analysis of source data is also shown in Supplementary Table 3. h–i, Rab11-GFP localization of 2nd instar larvae (65 h AEL) in the control (h) or upon DE-cad-RNAi (i). j–k, Confocal projections of phalloidin staining of the DT at mid-1st instar (42 h AEL) (left) or light microscopic views of aECM ridges at early L2 (around 50 h AEL) (right). Compared to the control (j), uif-RNAi (k) causes aECM ridge disorientation, which is especially evident at cell junctions. l, A single confocal section of DT stained for Uif (green) and E-cadherin (DE-cad, magenta) at 64 h AEL. In addition to the broad apical staining, Uif is preferentially detected along the longitudinal junctions (arrowhead) compared to the transverse junctions (arrow). In addition to strong junctional signals, DE-cad antibody weakly stains the cytoplasm³³. Scale bars: 20 μm. Images in a are representative of at least 10 larvae from ≥ 4 independent experiments. Data in b–g are aggregated from 2 independent experiments. Analysis of source data is also shown in Supplementary Table 3. Images in h and i are representative of at least 6 larvae from ≥ 2 independent experiments. Fluorescent images in j and k are representative of 6 larvae from ≥ 2 independent experiments. Light microscope views in j and k are representative of at least 10 larvae from ≥ 4 independent experiments. Images in l are representative of 6 larvae from 2 independent experiments.