Figure 8.
Abl and PTP61F oppositely regulate CME and crystal cell formation by modulating the phosphorylation status of Abi. (A) Single confocal sections of HmlΔ-GAL4/+, UAS-Abl/+; HmlΔ-GAL4/+ (HmlΔ > Abl), HmlΔ-GAL4/UAS-PTP61F (HmlΔ > PTP61F), UAS-Abl/+; HmlΔ-GAL4/UAS-PTP61F (HmlΔ > Abl, PTP61F), WT, Abl1/Abl4, PTP61FC05292/Df(3L)BSC289 (PTP61FP/PTP61FDf), and PTP61FC05292,Abl1/Df(3L)BSC289,Abl4 (PTP61FP,Abl1/PTP61FDf,Abl4) late-third instar larvae. Hemocytes were pulsed with Alexa Fluor 555-mBSA for 5 min, chased for 5 min, and stained with DAPI. (B) Quantification of mBSA intensity normalized to DAPI intensity in primary hemocytes of the indicated genotypes. Values are presented as percentages of HmlΔ-GAL4/+ or WT. n = 30 hemocytes. (C) Confocal images of peripheral hemocytes from late-third instar larvae of the indicated genotypes, stained with anti-Lz (green) and DAPI (blue). (D) Percentage of Lz+ crystal cells among all peripheral hemocytes (total DAPI count) in the indicated genotypes. n = 12 larvae. (E) Quantification of mBSA intensity normalized to DAPI intensity in primary hemocytes of the following genotypes: HmlΔ-GAL4/+, HmlΔ-GAL4/UAS-abi4YF (HmlΔ > abi4YF), HmlΔ-GAL4/UAS-abi4YE (HmlΔ > abi4YE), UAS-Abl/+; HmlΔ-GAL4/+ (HmlΔ > Abl), UAS-Abl/+; HmlΔ-GAL4/UAS-abi (HmlΔ > Abl,abi), UAS-Abl/+; HmlΔ-GAL4/UAS-abi4YF (HmlΔ > Abl,abiYF), UAS-Abl/+; HmlΔ-GAL4/UAS-abi4YE (HmlΔ > Abl,abiYE), WT, PTP61FC05292/Df(3L)BSC289 (PTP61FP/PTP61FDf), PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abi (PTP61FP/PTP61FDf, HmlΔ > abi), PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abiYF (PTP61FP/PTP61FDf, HmlΔ > abiYF), and PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abiYE (PTP61FP/PTP61FDf, HmlΔ > abiYE). n = 30 hemocytes. (F) Quantification of Lz+ crystal cells among all peripheral hemocytes (total DAPI count) in the indicated genotypes. n = 12 larvae. Data represent the mean ± SEM. Comparisons are with HmlΔ-GAL4/+ or WT unless otherwise indicated (***P < 0.001; ns, not significant). Statistical analyses were performed using a one-way ANOVA with the Tukey–Kramer post hoc test. Scale bars: 5 μm (A); 20 μm (C).

Abl and PTP61F oppositely regulate CME and crystal cell formation by modulating the phosphorylation status of Abi. (A) Single confocal sections of HmlΔ-GAL4/+, UAS-Abl/+; HmlΔ-GAL4/+ (HmlΔ > Abl), HmlΔ-GAL4/UAS-PTP61F (HmlΔ > PTP61F), UAS-Abl/+; HmlΔ-GAL4/UAS-PTP61F (HmlΔ > Abl, PTP61F), WT, Abl1/Abl4, PTP61FC05292/Df(3L)BSC289 (PTP61FP/PTP61FDf), and PTP61FC05292,Abl1/Df(3L)BSC289,Abl4 (PTP61FP,Abl1/PTP61FDf,Abl4) late-third instar larvae. Hemocytes were pulsed with Alexa Fluor 555-mBSA for 5 min, chased for 5 min, and stained with DAPI. (B) Quantification of mBSA intensity normalized to DAPI intensity in primary hemocytes of the indicated genotypes. Values are presented as percentages of HmlΔ-GAL4/+ or WT. n = 30 hemocytes. (C) Confocal images of peripheral hemocytes from late-third instar larvae of the indicated genotypes, stained with anti-Lz (green) and DAPI (blue). (D) Percentage of Lz+ crystal cells among all peripheral hemocytes (total DAPI count) in the indicated genotypes. n = 12 larvae. (E) Quantification of mBSA intensity normalized to DAPI intensity in primary hemocytes of the following genotypes: HmlΔ-GAL4/+, HmlΔ-GAL4/UAS-abi4YF (HmlΔ > abi4YF), HmlΔ-GAL4/UAS-abi4YE (HmlΔ > abi4YE), UAS-Abl/+; HmlΔ-GAL4/+ (HmlΔ > Abl), UAS-Abl/+; HmlΔ-GAL4/UAS-abi (HmlΔ > Abl,abi), UAS-Abl/+; HmlΔ-GAL4/UAS-abi4YF (HmlΔ > Abl,abiYF), UAS-Abl/+; HmlΔ-GAL4/UAS-abi4YE (HmlΔ > Abl,abiYE), WT, PTP61FC05292/Df(3L)BSC289 (PTP61FP/PTP61FDf), PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abi (PTP61FP/PTP61FDf, HmlΔ > abi), PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abiYF (PTP61FP/PTP61FDf, HmlΔ > abiYF), and PTP61FC05292,HmlΔ-GAL4/Df(3L)BSC289,UAS-abiYE (PTP61FP/PTP61FDf, HmlΔ > abiYE). n = 30 hemocytes. (F) Quantification of Lz+ crystal cells among all peripheral hemocytes (total DAPI count) in the indicated genotypes. n = 12 larvae. Data represent the mean ± SEM. Comparisons are with HmlΔ-GAL4/+ or WT unless otherwise indicated (***P < 0.001; ns, not significant). Statistical analyses were performed using a one-way ANOVA with the Tukey–Kramer post hoc test. Scale bars: 5 μm (A); 20 μm (C).

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