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Fig. 3 - Supplemental 1 Local manipulations with dominant-negative cdc42, rac1, or rhoab in peridermal keratinocytes. (A) Mosaic expression of krt4:TetGBDTRE-cdc42DN in krt4+ keratinocytes results in a significant reduction in cytoneme extension (F2, 289=50.82, p<0.0001, N=292 cells, 9 larvae total), and (E–E’) leads to local peridermal disorganization and elevated krtt1c19e expression in krt4+ keratinocytes compared to controls shown in D and E. (B, F–F’) Similar phenotypes are observed in krt4:TetGBDTRE-rac1DN expressing peridermal keratinocytes as in krt4:TetGBDTRE-cdc42DN expressing keratinocytes (F2,239=21.43, p<0.0001, N=242 cells, 9 larvae total). (C, G–G’) However, local expression of krt4:TetGBDTRE-rhoabDN shows no effects on cytoneme extension frequency and peridermal keratinocytes (F2,237=0.08942, p=0.9145, N=240 cells, 9 larvae total). Statistical significances were assessed by One-way ANOVA followed by Tukey’s HSD post hoc test. Scale bars: 20 µm. Error bars indicate mean ± SEM.