In vivo effects of kitlga and kitlgb at 72 hpf. (A) Representative photomicrographs of lcr:EGFP-transgenic reporter larvae at 72 hpf after injection of corresponding mRNAs. Control represents uninjected larvae. Images were acquired using a Zeiss Axio Zoom.V16 with Zeiss Axiocam 506 mono microscope camera and ZEN (blue edition) software. (B) Quantification of lcr:EGFP⁺ cells in embryonic tails from fish in panel A. Values were plotted relative to the mean of epo (relative expression level 1). (C) FACS analysis of lcr:EGFP⁺ cells in whole embryos. (D) Representative photomicrographs of benzidine-stained embryonic tails at 72 hpf after injection of corresponding mRNAs. Control represents uninjected embryos. Note the increase in pigmentation in the dorsal part of kitlga-injected embryos. Z-stacks were acquired using a Zeiss Axio Zoom.V16 with aa Zeiss Axiocam 105 color camera (total magnification ×125). Images were processed using the Extended Depth of Focus module in ZEN (blue edition) software. (E) Quantification of the benzidine-stained embryonic tails in (D). Values were plotted relative to the mean of epo (relative expression level 1). (F) qPCR expression analysis of gata1a and hbbe1 from whole injected embryos at 72 hpf. Data were normalized using mob4 as the housekeeping gene and plotted relative to the mean of uninjected control (relative expression level 1). Bars represent a mean of 3 triplicates with standard deviation. *P ≤ .05, **P ≤ .01, ***P ≤ .001, ****P ≤ .0001, standard 1-way analysis of variance. n.s., not significant (P > .05).