Fig. 8: Ihog expression focalizes Cheerio at the plasma membrane.

Α–a’ Cher^YFP (green) wing disc expressing Ihog^RFP (red) at the dorsal compartment, keeping the ventral half as control. To analyse the co-localization of Ihog^RFP and Cher^YFP, the samples were immunostained with an α-GFP antibody. A1 Cher^YFP in grey. a, a’). Enlarged insets showing Cher^YFP expression. Note the increased focalization of the Cher signal to spots at the plasma membrane upon overexpression of Ihog^RFP (a’). B–b’ Cher^YFP (green) wing disc expressing UAS.IhogΔC^RFP (red) at the dorsal half, keeping the ventral half as control. B1 Cher^YFP in grey. Note the focalization effect over Cher is lost when Ihog lacks the C-terminal region (b, b’). C Representation of the Cher^YFP signal in A and B within an area in normal conditions (orange), after UAS.Ihog^RFP (blue) or after UAS.IhogΔC^RFP expression (grey). Note the Cher^YFP signal after Ihog^RFP expression presents picks of punctual signal more intense than after IhogΔC^RFP expression. D Quantification of the Cher signal distribution as a ratio between the signal standard deviation from equal areas within the dorsal and ventral halves (control) to normalize (wing discs per treatment for WT n = 7, for Ihog n = 5, for IhogΔC^RFP n = 7). Statistical analysis showed the rate values for Cher signal distribution when UAS.Ihog^RFP (orange) is expressed (p < 0.01) and no difference with wild type (green) or after expressing UAS.IhogΔC^RFP (blue) (p = 1). E–G Cher^YFP salivary gland cells showing Cher protein relocalization by Ihog^RFP (red) overexpression. Cher^YFP channel (green, grey) shows a drastic focalization of the Cher signal to spots at the plasma membrane upon overexpression of Ihog^RFP (F1 compared to E). G Graphical representation of (F) under normal conditions (blue) and after Ihog^RFP overexpression (red). Note the increase in signal levels and the higher picks indicating Cher focalization to small segments within the membrane. H Basal section of a Cher^YFP wing disc after UAS.EGFR^RNAi and UAS.RHGmiRNA in the dorsal side. I Quantifications analysed by two sided pairwise comparison with Wilcoxon test show the downregulation of Cher^YFP in the absence of EGFR (p < 0.0001). Scale bars = 10 μm, enlarged inserts 5 μm. Source data are provided as a Source Data file.