Figure 4
Figure 4. Cell cycle profile of primitive myeloid cells in irf8 MOsp morphants. (A-B) BrdU and GFP double staining indicates the neutrophils in S phase in the CHT region of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (C-D) pH3 and GFP double staining reveals the neutrophils in M phase in the CHT region of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (E) Histogram represents the percentage of BrdU+mpx-GFP+ (S phase) and pH3+mpx-GFP+ (M phase) cells in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). *Statistical differences with corresponding control (t test, P < .05). (F-G) BrdU and Lcp1 double staining shows the overall myeloid cells in S phase in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (H-I) pH3 and Lcp1 double staining shows the overall myeloid cells in M phase in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (J) Histogram reveals the percentage of BrdU+Lcp1+ (S phase) and pH3+/Lcp1+ (M phase) cells in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). (K-L) GFP and Lcp1 double staining presents neutrophils in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (M) Histogram shows the percentage of mpx-GFP+Lcp1+ cells (neutrophils) in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). *Statistical differences with corresponding control (t test, P < .05). White arrows indicate the costaining of BrdU and GFP in panels A and B, pH3 and GFP in panels C and D, BrdU and Lcp1 in panels F and G, pH3 and Lcp1 in panels H and I, and GFP and Lcp1 in panels K and L. (N-O) WISH indicates lcp1 RNA expression in 32 hpf control embryo (n = 35/35) and irf8 MOsp morphant (n = 29/29). Blue arrows represent the WISH signals. (P) Histogram reveals the overall myeloid cell number positive for lcp1 in 32 hpf control embryos and irf8 MOsp morphants (n ≥ 10, mean ± SE).

Cell cycle profile of primitive myeloid cells in irf8 MOsp morphants. (A-B) BrdU and GFP double staining indicates the neutrophils in S phase in the CHT region of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (C-D) pH3 and GFP double staining reveals the neutrophils in M phase in the CHT region of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (E) Histogram represents the percentage of BrdU+mpx-GFP+ (S phase) and pH3+mpx-GFP+ (M phase) cells in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). *Statistical differences with corresponding control (t test, P < .05). (F-G) BrdU and Lcp1 double staining shows the overall myeloid cells in S phase in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (H-I) pH3 and Lcp1 double staining shows the overall myeloid cells in M phase in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (J) Histogram reveals the percentage of BrdU+Lcp1+ (S phase) and pH3+/Lcp1+ (M phase) cells in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). (K-L) GFP and Lcp1 double staining presents neutrophils in the CHT of 32 hpf Tg(mpx:eGFP) control embryo (n = 8/8) and irf8 MOsp morphant (n = 7/7). (M) Histogram shows the percentage of mpx-GFP+Lcp1+ cells (neutrophils) in 32 hpf Tg(mpx:eGFP) control embryos and irf8 MOsp morphants (n ≥ 7, mean ± SE). *Statistical differences with corresponding control (t test, P < .05). White arrows indicate the costaining of BrdU and GFP in panels A and B, pH3 and GFP in panels C and D, BrdU and Lcp1 in panels F and G, pH3 and Lcp1 in panels H and I, and GFP and Lcp1 in panels K and L. (N-O) WISH indicates lcp1 RNA expression in 32 hpf control embryo (n = 35/35) and irf8 MOsp morphant (n = 29/29). Blue arrows represent the WISH signals. (P) Histogram reveals the overall myeloid cell number positive for lcp1 in 32 hpf control embryos and irf8 MOsp morphants (n ≥ 10, mean ± SE).

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