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. 2020 Jan 15;295(8):2421–2437. doi: 10.1074/jbc.RA119.009451

Figure 4.

Figure 4.

Dorsal is activated in dElys depletion and shows induced apoptotic response. A, detection of Dorsal with anti-Dorsal (green) and dElys antibodies (red) in control (upper panels) and dElys RNAi (ubiquitous Act5C–GAL4 driven, lower panels) third instar larval salivary gland nuclei. DNA is stained with DAPI. Scale bar, 5 μm. Control is WT flies crossed with the Act5C–GAL4 driver. B, detection of Cactus in third instar larval salivary gland cells with anti-Cactus (green) as mentioned in A. Scale bar, 5 μm. C, detection of apoptosis in third instar larval salivary gland cells with anti-Drosophila caspase-1 antibody (red) and FG-nucleoporins by mAb414 antibodies in control (upper panels) and dElys depletion (ubiquitous Act5C–GAL4 driven, lower panels). DNA is stained with DAPI. Scale bar, 5 μm. Control is WT flies crossed with the Act5C–GAL4 driver. D–F, quantification of intensities of indicated molecules in control and dElys RNAi condition. The intensity of each molecule was normalized to the intensity of DAPI. Data are represented from at least three independent experiments. At least 45 nuclei were analyzed from 7 to 8 pairs of salivary glands. Statistical significance was derived from the Student's t test. Error bars represent S.E. *** represents p < 0.0001. G, expression analysis of apoptosis regulatory genes, reaper, hid, and diap-1 by quantitative PCR in cDNA from third instar larval head complex of control and dElys RNAi (ubiquitous Act5C–GAL4 driven). Data are represented from at least three independent experiments. Statistical significance was derived from the Student's t test. The error bars represent the standard deviation. *** represents p < 0.0001, and ** represents p < 0.001. H, expressions of Dorsal target genes snail, twist, rho, dpp, short gastrulation (sog), and drosomycin analyzed by quantitative PCR in control and dElys RNAi (ubiquitous Act5C–GAL4 driven) third instar larval head complex lysate. Data are represented from at least three independent experiments. Statistical significance was derived from the Student's t test. The error bars represent the standard deviation. *** represents p < 0.0001, and ** represents p < 0.001, and * represents p < 0.05; ns is nonsignificant.