Fig. 6.

Sustained LIX1 expression decreases YAP1 activity and hinders smooth muscle cell differentiation. a Serial transverse sections of E7 stomachs analysed either by in situ hybridization using the LIX1 riboprobe or by immunofluorescence with anti-αSMA and anti-CALPONIN antibodies. Nuclei were visualized with Hoechst. Black arrowheads in the LIX1 panels indicate the area imaged at high power in the αSMA and CALPONIN panels. b RT-qPCR analysis of relative mRNA expression in E7 stomachs. Data were normalized to GAPDH expression. Values are presented as the mean ± standard deviation (SD) of n = 7 controls vs. n = 6 LIX1-expressing stomachs. *P < 0.05; **P < 0.01 by one-tailed Mann–Whitney test. c Serial transverse sections of E7 stomachs analysed either by in situ hybridization using the LIX1 riboprobe or by immunofluorescence with anti-PH3 antibodies. Black arrowheads in the LIX1 panels indicate the area imaged at high power in the PH3 panels. Nuclei were visualized with Hoechst. Scale bars, 500 μm. Graphs represent the quantification of PH3-positive cells and cell density. Normalized expression levels were converted to fold changes. Values are presented as the mean ± standard error of the mean of n = 16 controls vs. n = 16 LIX1-expressing stomachs. ****P < 0.0001 by two-tailed Mann–Whitney tests. ns, not significant. d Western blot analysis of YAP and phospho-YAP (72 kDa) protein levels. Equal loading was verified by GAPDH expression (37 kDa). Graph represents the quantification of western blot data. Values are presented as the mean ± SD of n = 4 controls vs. n = 4 LIX1-expressing stomachs. *P < 0.05 by one-tailed Mann–Whitney test. Raw data are shown in Additional file 12. e RT-qPCR analysis of relative mRNA expression in E7 stomachs. Data were normalized to GAPDH expression. Normalized expression levels were converted to fold changes. Values are presented as the mean ± SD of n = 7 controls vs. n = 6 LIX1-expressing stomachs. *P < 0.05; **P < 0.01 by one-tailed Mann–Whitney tests