Extended Data Figure 6: p5325,26,53,54/+ embryo tissues display increased apoptosis and decreased proliferation. | Nature

Extended Data Figure 6: p5325,26,53,54/+ embryo tissues display increased apoptosis and decreased proliferation.

From: Inappropriate p53 activation during development induces features of CHARGE syndrome

Extended Data Figure 6

a, Left, Immunofluorescence for phospho-histone H3 (red) in the retina of E13.5 control and p5325,26,53,54/+ embryos. Right, Quantification of phospho-H3-positive cells per retina area relative to littermate controls. **, P = 0.006 by one-tailed Welch’s t-test (n = 4). Original magnification, ×200. b, Left, Immunohistochemistry for cleaved caspase 3 (CC3) in thymi of control (left) and p5325,26,53,54/+ (right) embryos. Inset, Magnified image of CC3-positive region. Right, Quantification of CC3-positive cells per thymic area. *, P = 0.02 by one-tailed Student’s t-test (n = 4). Original magnification, ×400. c, Immunofluorescence for PAX3 (green) in NCCs of E9.5 control and p5325,26,53,54/+ embryos was used to identify NCCs in Fig. 2f. Original magnification, ×200. d, Left, Immunofluorescence for CC3 (red) and PAX3 (green) in NCCs of E9.5 control and p5325,26,53,54/+ embryos. p5325,26,53,54/+ embryos have more apoptotic (red) NCCs, as determined by PAX3-positive staining (green), than littermate controls. Right, Quantification of CC3-positive cells per total NCC number. P = 0.14 by one-tailed Student’s t-test (n = 4). Original magnification, ×200. e, Left, Immunofluorescence for CC3 (red) in the otic vesicles of E9.5 control and p5325,26,53,54/+ embryos. Right, Quantification of CC3-positive cells per total cell number. *, P = 0.03 by one-tailed Student’s t-test (n = 3). Original magnification, ×200. f, CC3 staining in whole-mount E8.5 control and p5325,26,53,54/+ embryos reveals enhanced apoptosis in the neuroepithelium of p5325,26,53,54/+ embryos (right) but not in controls (left). The close-up shows a magnification of the caudal neuroepithelium (bottom). Arrows indicate CC3-positive regions. Original magnification top, ×50.

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