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Figure 5
Selective ablation of BLECs decreased internalization of mA?42 and pronephric accumulation. (A) A schematic diagram of the experimental setting. BLEC-specific ablation using the confocal laser. (B,C) Confocal images of BLECs in the double transgenic Tg(mrc1a:mCherry); Tg(kdrl:EGFP) at 3 dpf before laser irradiation (B) and after ablation (C). Yellow arrows indicate the ablated BLECs. Scale bars = 50 ?m. (D) Confocal images of mrc1a+ BLECs in the loop of the optic before ablation (D) and after ablation and mA?42 injection (D??D?). (D?) shows red channel, (D?) shows mA?42 (HiLyte647, white) and (D?) is a merged image. Dotted lines denote the loop of BLECs. (E) Confocal images of mrc1a+ BLECs with mA?42 injection (non-ablated control). Scale bars = 50 ?m. (E?) shows mA?42 (HiLyte647, white) and (E??) shows merged images of BLECs and mA?42. (F) Quantification of the relative intensity ratio between the pronephros and hindbrain. Data are presented as mean ± SEM. Statistical significance was determined by two-tailed unpaired t-test. n = 15 for non-ablated control, n = 8 for ablated. p = 0.0092. (G) Quantification of area fraction (%) occupied by mrc1a+ BLECs. Data are presented as mean ± SEM. Statistical significance was determined by ordinary one-way ANOVA with Tukey?s test. **, p < 0.01; ****, p < 0.0001.