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. 2022 Jan 13;11:e61389. doi: 10.7554/eLife.61389

Figure 2. p53A is necessary and sufficient for IR-induced apoptosis in the soma.

(A–B) The p53 isoform-specific mutants created at the endogenous p53 locus with CRISPR / Cas9. Each allele is a small deletion (red asterisk) in the unique 5’ coding exon of p53A (A) and p53B (B) mRNAs. The p53A2.3 (A-B+) mutant impairs expression of isoforms p53A and p53C (gray shading) but not p53B, whereas the p53B41.5 (A+B-) mutant eliminates expression of p53B (gray shading) but not p53A (see Figure 2—figure supplement 1). (C–H) Apoptotic response to IR of stage six somatic follicle cells, assayed by TUNEL (red), with DNA stained with DAPI (gray). (C–D) TUNEL-labeled follicle cells from a p53+ (A+B+) wild type female without (C) or 4 hr after IR (D). (E–G) TUNEL labeling of follicle cells after IR from p535A-1-4 (A-B-) null (E), p53A2.3 (A-B+) (F), and p53B41.5 (A+B-) (G) mutant females. Scale bars are 10 μm. (H) Quantification of the average number of TUNEL-labeled follicle cells in stage six egg chambers for the genotypes and treatments shown in (C–G). Averages are based on 10 egg chambers per genotype with two biological replicates. Error bars are S.E.M. ****: p < 0.0001 by unpaired Student’s t test.

Figure 2—source data 1. Counts of TUNEL-positive follicle cells for Figure 2.

Figure 2.

Figure 2—figure supplement 1. p53 isoform-specific alleles.

Figure 2—figure supplement 1.

(A) Left: Shown are the location of p53A2.3 and p53B41.5 deletion alleles and the coordinates of primers (red font) used for RT-PCR. Expanded maps below indicate the coordinates of the deletion endpoints within the p53A and p53B mRNA isoforms (dotted lines and straight arrows). Right: DNA Sequence of p53A2.3 and p53B41.5 deletion alleles (top) aligned with Drosophila reference genome sequence (WT, bottom). Red nucleotides indicate unique nucleotide insertions. All genomic coordinates are based on Drosophila melanogaster genome release 6.32. (B, B’) RT-PCR with the primer pairs indicated in (A) that are specific to p53A mRNA (B) or p53B mRNA (B’) from p53+ (A+B+), p53A2.3 (A-B+), p53B41.5 (A+B-), p535A-1-4 (A-B-). Black arrows indicate position of p53A and p53B PCR products.
Figure 2—figure supplement 1—source data 1. Counts of TUNEL-positive follicle cells for Figure 2.