International Journal of Stem Cells : eISSN 2005-5447

Fig. 6.

Download original image
Fig. 6. Enhanced mesendodermal differentiation mediated by Wnt signaling pathway partially accounted for the effects of p53 on the cardiac differentiation from hiPSC. (A) The qRT-PCR results showing hiPSC with lenti-P53 had raised mRNA level of WNT3. It was further confirmed by representative image of Western Blot (B) and quantification by densitometry (C). (D) The mRNA level of WNT3 in hiPSC was decreased by knockdown of P53 as shown by qRT-PCR results. (E) Representative image of Western Blot displayed notably reduced protein level of WNT3 caused by knockdown of P53, and quantification was done by densitometry (G∼I). WNT3 gene was successfully downregulated by Sh-WNT3 using qRT-PCR (G) and Western Blot analysis (H, I). (J, K) The qRT-PCR results showed that Sh-WNT3 notably reversed P53-caused upregulation of mesendodermal markers (BRACHYURY and EOMES) (J) and cardiac mesodermal makers (MESP1 and KDR) (K) at day 3 and 5 from differentiation respectively. (L, M) The qRT-PCR results showed that knockdown of WNT3 significantly abrogated P53-dependent increased mRNA levels of mesendodermal markers (BRACHYURY and EOMES) at day 3 from differentiation (L) and cardiac mesodermal makers (MESP1 and KDR) at day 5 (M). Expression values of the PCR analysis were normalized to GAPDH. Data are presented as mean±SD (t test, *p<0.05; n=3, performed in 3 independent experiments).
International Journal of Stem Cells 2021;14:410-22 https://doi.org/10.15283/ijsc21051
© 2021 International Journal of Stem Cells