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Fig. 3 Knockdown of cited3 results in increased cell death but it does not affect proliferation.
Embryos that are injected with the control MO (A–C), cited3 MO (D–F), cited3 MO + cited3 RNA (G–I) were fixed at 2 dpf and immunostained with anti-Mef2 antibody (A,D,G) and TMR red in situ cell death detection kit (B,E,H). Tunel positive cells were counted in non-muscle cells as well as in muscle cells in each group of embryos from 50–100 sections obtained from trunk and tail somites. More apoptotic muscle cells were detected in embryos that are injected with the cited3 MO and fixed at 2 dpf and 3 dpf (M). Values were normalized compared to cited3 morphant values. Similarly, embryos injected with the control MO (J), cited3 MO (K), cited3 MO + cited3 RNA (L) was pulsed with BrdU from 30 hpf to 42 hpf, and were fixed and immunostained with anti-BrdU and MF20 antibodies (images of 16th–18th somites). There is no significant difference between the three treated groups (J,K,L) when the BrdU-positive muscle cells were counted (data not shown). Scale bars: 25 μm in A and 50 μm in K.