Fig. 2: ptip−/− and oncogenic Ras cooperatively induce nonautonomous tissue overgrowth via wild-type p53. | Communications Biology

Fig. 2: ptip−/− and oncogenic Ras cooperatively induce nonautonomous tissue overgrowth via wild-type p53.

From: Cooperation between oncogenic Ras and wild-type p53 stimulates STAT non-cell autonomously to promote tumor radioresistance

Fig. 2

(a–i′) Representative images of dissected eye imaginal discs containing ptip−/− or RasV12 or RasV12ptip−/− double mutant clones (GFP) stained with DAPI to detect DNA or anti-phosphorylated H2AV antibodies to detect DNA damage (a–c’) or anti-p53 (d-e′) or anti-dacapo (dap/p21) (f–g′) or anti-phosphorylated JNK (h-i′) antibodies to detect cellular response to DNA damage. Scale bars are 20 µm. (j, k) Quantitative Polymerase Chain Reaction (qPCR) data showing expression of p53 or dap/p21 in wild type versus ptip−/− eye imaginal discs (j) or the expression of p53 in RasV12 or ptip−/− or RasV12ptip−/− eye imaginal discs (k). Expression was normalized to the transcript abundance of the housekeeping gene rp49. Error bars denote standard deviation (SD) values. P values are derived from Student’s t test analyses. (l–r) Matched light and fluorescence images of adult eyes containing GFP-labeled clones. The respective clone genotypes are indicated at the top of each panel. The corresponding fluorescent images are shown below in (l′–r′). GPF-negative tissues represent wild-type tissues. Scale bars are 150 µm. (s) Quantification of the nonautonomous growth phenotype of adult eyes containing clones of the indicated genotypes: RasV12ptip−/−, RasV12ptip−/−BskDN, RasV12ptip−/−p53R155H, or RasV12ptip−/−p53RNAi. (t–u″) Genetic juxtaposition of GFP-labeled RasV12 clones with RFP-labeled RasV12 clones (t-t″, controls) or with RFP-labeled clones of cells coexpressing RasV12 and wild-type p53 (RasV12, p53OE) (u-u″). GFP-positive RasV12 clones are surrounded by RFP/GFP double-positive RasV12 clones (t-t″) or by RFP/GFP double-positive RasV12, p53OE clones (u-u″). Brain cephalic complex images showing the growth of RasV12 clones when juxtaposed to RasV12 or to RasV12, p53OE clones are shown in t and u, respectively. Dotted white lines (t′, t″, u′, u″) represent tissue boundaries. Scale bars are 100 µm. (v) Quantification of eye tissue sizes from (t–u″). Sample size N = 10 tissues per genotype. Error bars denote standard error of the mean (SEM) values. P values are derived from Student’s t test analyses. Effect size (Cohen’s d values) for (j), (k), and (v) is greater than 0.8.

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