Fig. 3
Btk29A mutations suppress the Ptp4EPtp10D phenotypes. a Confocal frames showing the dorsal trunk (DT) of living wild-type, Ptp4EPtp10D, Btk29A5610 and Ptp4EPtp10D;Btk29A5610 mutant embryos expressing btl>ANF-GFP (grey). The indicated time points represent hours after egg laying (AEL). The premature ANF-GFP luminal clearance in Ptp4EPtp10D (arrowheads) is repressed in Ptp4EPtp10D;Btk29A5610 triple mutant embryos. b Schematics of the luminal clearance in wild-type, Ptp4EPtp10D, Btk29A5610 and Ptp4EPtp10D;Btk29A5610 embryos. The time interval of luminal uptake in wild-type embryos is indicated (green box). c Graph showing the average number of wild-type (n = 39), Ptp4EPtp10D (n = 51) and Ptp4EPtp10D;Btk29A5610 (n = 65) embryos with luminal ANF-GFP at 17.5 h AEL (1.5 h prior to the luminal clearance initiation in wild-type). ***P < 0.0001 (unpaired two tailed t test). d Confocal images of the DT in wild-type, Ptp4EPtp10D, Btk29A5610 and Ptp4EPtp10D;Btk29A5610 mutant embryos (stage 17) stained for the luminal protein GASP (magenta in upper row panels) or the adherens junction protein DE-Cadherin (grey in lower row panels). e Plots depicting the average diameter of the DT (Tr8) in wild-type (n = 6), Btk29A5610 (n = 8), Ptp4EPtp10D (n = 7), Ptp4EPtp10D;Btk29A5610 (n = 8) and Ptp4EPtp10D;Df235/Btk29A5610 (n = 6) embryos (stage 17). The boxplot shows the median (internal line) and the range of the data is from 25th to 75th percentile. The bars present maxima and minima values. Unpaired two tailed t tests were performed in comparison to Ptp4EPtp10D data set. **P < 0.006. f Selected frames from live imaging of the tracheal DT of btl>Btk29A and btl>Btk29AKD embryos expressing ANF-GFP (grey). g Plots showing the percentage of wild-type (n = 14), btl>Btk29A (Type 2 or long isoform) (n = 28) and btl>Btk29AKD (n = 32) embryos that completed ANF-GFP luminal clearance. Unpaired two tailed t test was performed to compare btl>Btk29A and btl>Btk29AKD data sets. ***P < 0.0001. h Plot showing the percentage of wild-type (n = 54), Btk29A5610 (n = 67), Df235/Btk29A5610 (n = 75), Btk29A5610;btl>Btk29A-1 (type-1) (n = 58) and Btk29A5610;btl>Btk29A-2 (type-2) (n = 72) embryos that complete luminal ANF-GFP clearance. *P < 0.01, **P < 0.001, (unpaired two tailed t tests). Error bars show s.e.m. Scale bars, 10 μm
