Figure 4
Figure 4. Functional analysis of gcsfr morphants. (A) Quantitation of TUNEL assay performed on 22-hpf embryos previously injected with either scramMo or gcsfrMo1. Only apoptotic cells on the yolk were counted, with the mean (dashed red line) and 95% confidence interval (red lines) indicated. (B) Fluorescence analysis of lyz::EGFP cells from 96-hpf embryos based on EGFP fluorescence, forward scatter (FSC), and side scatter (SSC) characteristics. (C) RT-PCR of various genes from total RNA extracted from lyz::EGFP-expressing fluorescence-activated cell sorting (FACS)–isolated cells. (D-I) In vivo visualization of lyz::EGFP cells at the developmental time points indicated in embryos previously injected with scramMo (D-F) or gcsfrMo1 (G-I). (J-M) Wounding-induced migration of lyz::EGFP cells in 72-hpf embryos that had been previously injected with scramMo (J,K) and gcsfrMo1 (L,M) at 15 minutes and 105 minutes after injury. (N-S) Phagocytosis assay of lyz::EGFP cells in 48-hpf embryos injected with pHrodo E coli BioParticle conjugate that had been previously injected with scramMo (N-P) or gcsfrMo1 (Q-S). Panels show separate EGFP (N,Q), rhodamine (O,R), and merged (P,S) images. (T,U) Three-dimensional isosurface reconstruction of lyz::EGFP cells on the yolk surface of 20-hpf embryos previously injected with scramMo (T) or gcsfrMo1 (U). (V,W) Myeloperoxidase staining of 96-hpf embryos that had been previously injected with scramMo (V) or gcsfrMo1 (W).

Functional analysis of gcsfr morphants. (A) Quantitation of TUNEL assay performed on 22-hpf embryos previously injected with either scramMo or gcsfrMo1. Only apoptotic cells on the yolk were counted, with the mean (dashed red line) and 95% confidence interval (red lines) indicated. (B) Fluorescence analysis of lyz::EGFP cells from 96-hpf embryos based on EGFP fluorescence, forward scatter (FSC), and side scatter (SSC) characteristics. (C) RT-PCR of various genes from total RNA extracted from lyz::EGFP-expressing fluorescence-activated cell sorting (FACS)–isolated cells. (D-I) In vivo visualization of lyz::EGFP cells at the developmental time points indicated in embryos previously injected with scramMo (D-F) or gcsfrMo1 (G-I). (J-M) Wounding-induced migration of lyz::EGFP cells in 72-hpf embryos that had been previously injected with scramMo (J,K) and gcsfrMo1 (L,M) at 15 minutes and 105 minutes after injury. (N-S) Phagocytosis assay of lyz::EGFP cells in 48-hpf embryos injected with pHrodo E coli BioParticle conjugate that had been previously injected with scramMo (N-P) or gcsfrMo1 (Q-S). Panels show separate EGFP (N,Q), rhodamine (O,R), and merged (P,S) images. (T,U) Three-dimensional isosurface reconstruction of lyz::EGFP cells on the yolk surface of 20-hpf embryos previously injected with scramMo (T) or gcsfrMo1 (U). (V,W) Myeloperoxidase staining of 96-hpf embryos that had been previously injected with scramMo (V) or gcsfrMo1 (W).

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