Journal: Birth Defects Research
Article Title: Pharmacological Inhibition of the Spliceosome SF3b Complex by Pladienolide‐B Elicits Craniofacial Developmental Defects in Mouse and Zebrafish
doi: 10.1002/bdr2.2404
Figure Lengend Snippet: Phenotype of Sf3B4 +/− embryos exposed to PB. (a) Ratio of phenotype of wild‐type (top, n = 11) and Sf3B4 +/− (bottom, n = 6) embryos at E10.5 exposed to PB at E8.25. Embryos were obtained from three pregnant mice. (b) Wild‐type and (c) Sf3B4 +/− embryos at E9.5 exposed to PB at E8.25. Yellow arrow indicates the PA1 in the Sf3B4 +/− embryo. (d–k) Frontal sections of E8.5 wild‐type (d–g) and Sf3B4 +/− (h–k) embryos exposed to PB at E8.25. (d, f, h, j) Hoechest, (e, i) PHH3, and (g, k) CC3 fluorescent signals. (l) Ratio of PHH3 and CC3 positive cells in the neuroepithelium and mesenchyme in E8.5 wild‐type and Sf3B4 +/− embryos exposed to PB at E8.25. (m–u) Frontal sections of wild‐type (m–p) and Sf3B4 +/− (q–u) embryos at E9.5 exposed to PB at E8.25. (m, o, q, s) Hoechest, (n, r) PHH3, and (p, t) CC3 fluorescent signals. (u) Fluorescent signal of AP2 on the same section as (t). (v) Overlay image of the white box area in (s–u). Cells overlapping CC3 (green) and AP2 (magenta) signals are indicated in white. n = 3 for each experiment in (d–k) and (m–v). (w) Ratio of PHH3 and CC3 positive cells in the neuroepithelium and mesenchyme in E9.5 wild‐type and Sf3B4 +/− embryos exposed to PB at E8.25. For (l) and (w), the cell rates were calculated by the positive cell number per nucleus number in the same area. The dots in (l) and (w) represent individual embryos obtained from at least two pregnant mice. *: p < 0.05, ***: p < 0.001 (Student's t ‐test). Scale bar: 400 μm for (b, c), 50 μm for (d, h) and 100 μm for (m, q).
Article Snippet: The sections were washed by PBS and treated with 5% goat or horse serum for 30 min and then incubated with the primary antibody; N‐cadherin (BD Transduction, #610920, 1:200), Phospho‐Histone H3 (Cell Signaling Technology, #9701, 1:200), Cleaved Caspase‐3 (Cell Signaling Technology, #9661, 1:200), or AP2 (DSHB, #3B‐5, 1:100) overnight.
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