Fig. 4: The 20HE-Crol-Wg axis in EC controls Wnt/Wg signalling activity in ISC/EB.
From: Steroid hormone-induced wingless ligands tune female intestinal size in Drosophila
A Schematic of the system enabling EC manipulation and simultaneous visualization of Wnt/Wg activity in ISC/EB. EC specificity of mex-Gal4 allows manipulation of UAS-driven transgenes that are timely controlled by the ubiquitously expressed temperature sensitive Gal4 repressor (tub-Gal80ts) and combined with the fz3-RFP reporter reflecting Wnt/Wg signalling activity in ISC/EB independent of Gal4. B–D fz3-RFP intensities in ISC/EB with (B) OE and KD of EcI, crol (F003414, BL41669) and wg (>wg-RNAi: p = 0.0005), (C) EC specific OE of crol (F003414) combined with >wg-RNAi, and (D) ectopic expression of >hZNF267 in crol depleted EC (BL41669). B–D Experiments have been performed in parallel. E-M’ Confocal images of fz3-RFP combined with mex> and tub-Gal80ts from R5 regions of MF midguts after 24 h in (E–E’) controls, EC specific OE of (F–F’) EcI, (G–G’) crol (H–H’) with simultaneous KD of wg, and (I–I’) wg, and KD of (J–J’) EcI, (K–K’) crol (L–L’) with simultaneous expression of >hZNF267, and (M–M’) KD of wg in EC. E–M Sole fz3-RFP signal is shown in greyscale and (E’–M’) in red colour combined with αArm and αPros staining for identification of ISC/EB. Exemplary measured ISC/EB are marked by white arrows. Scale bar is 20 µm. N–P’ MF midguts after 3 d with wg::GFP crossed to (N–N’) mex > ,tub-Gal80ts serving as control, (O–O’) combined with EC specific OE of crol and (P–P’) >crol-RNAi. N–P Sole Wg::GFP signal is shown in greyscale and (N’–P’) in green colour combined with αArm and αPros staining for identification of ISC/EB, EE and EC. Exemplary measured EC are outlined by red dashed lines. Scale bar is 20 µm. Q Wg::GFP levels in EC upon mex> driven manipulations of crol (F003414, BL41669). B–D, Q Scatter dot plots show individual values with indication of means ± SD. N and n values represent number of cells and number of biological replicas, respectively. Asterisks denote significances from multiple comparisons by Kruskal-Wallis tests (***p < 0.001;****p < 0.0001). Source data are provided as a Source Data file.
