Extended Data Fig. 10: Characterization of the effects of aPKC Trim-Away in mouse embryos.
From: Initiation of a conserved trophectoderm program in human, cow and mouse embryos
a, Immunofluorescence analysis of E-cadherin, mCherry and DAPI nuclear staining in mouse morula embryos in control and Trim-Away conditions (n = 10). b, Immunofluorescence analysis of anti-mouse IgG secondary antibody to detect the electroporated mouse anti-aPKC primary antibody, E-cadherin, rabbit anti-aPKC primary antibody to detect the aPKC protein and DAPI nuclear staining in mouse morula embryos in control and Trim-Away conditions. c, Quantification of aPKC cortical intensity (based on rabbit anti-aPKC signal) in outer cells at the morula stage of mouse embryos shown in b (n = 270 cells from 30 embryos). Data are presented as mean ± s.d. Two-tailed Student’s t-test, ****P < 0.0001. d, Immunofluorescence analysis of GATA3, YAP1, anti-mouse IgG secondary antibody to detect the electroporated mouse anti-aPKC primary antibody and DAPI nuclear staining in mouse morula embryos in control and Trim-Away conditions. Yellow arrowheads point to decreased YAP1 and GATA3 expression. e, f, Quantification of YAP1 (e) and GATA3 (f) normalized fluorescence intensity in outer cells of mouse embryos shown in d (n = 281 cells for YAP1 from 32 embryos, n = 263 cells for GATA3 from 31 embryos). Data are presented as mean ± s.d. Two-tailed Student’s t-test, ***P = 0.00078 for GATA3 distribution, ****P < 0.0001 for YAP1 distribution. g, Immunofluorescence analysis of SOX2, rabbit anti-aPKC primary antibody to detect the aPKC protein, anti-mouse IgG secondary antibody to detect the electroporated mouse anti-aPKC primary antibody and DAPI nuclear staining in mouse morula embryos in control and Trim-Away conditions. Yellow arrows point to outer cells expressing SOX2 in a mouse morula embryo. n = 2 independent experiments. h, Quantification of SOX2 normalized fluorescence intensity in outer cells of mouse embryos shown in g (n = 120 cells 18 embryos). Data are presented as mean ± s.d. Two-tailed Student’s t-test, *P = 0.026. i, Immunofluorescence analysis of TEAD4, anti-mouse IgG secondary antibody to detect the electroporated mouse anti-aPKC primary antibody and DAPI nuclear staining in mouse morula embryos in control and Trim-Away conditions. n = 2 independent experiments. j, Quantification of TEAD4 normalized fluorescence intensity in outer cells of mouse embryos shown in i (n = 90 cells from 10 embryos). Data are presented as mean ± s.d. Two-tailed Student’s t-test, ns, not significant. Scale bars, 25 μm. More details about statistics and reproducibility are provided in Methods.
