Fig 5. In utero pifithrin-α treatment improves colon colonization by TashTTg/Tg ENCCs in a male-specific manner.
(A) Representative confocal images of YFP-labeled ENCCs in the colon of male and female e15.5 TashTTg/Tg embryos that were exposed to either pifithrin-α or only vehicle (DMSO). The white arrow indicates the migration front of vagal-derived ENCCs. Scale bar, 500 μm. (B) Sex-stratified quantitative analysis of vagal-derived ENCC colonization (in % of total colon length) using images such as those displayed in A. (C-D) RT-qPCR analysis of endogenous mRNA expression of Ddx3y (C) and Ddx3x (D) genes in TashTTg/Tg ENCCs recovered by FACS from male embryos exposed to pifithrin-α or only vehicle (DMSO), either during (e12.5) or after (e15.5) the treatment period. (E) Correlation between Ddx3y expression levels in ENCCs and extent of ENCC colonization in the colon for e15.5 male TashTTg/Tg embryos that were exposed to pifithrin-α. (R, Pearson’s correlation coefficient; *P≤0.05; *** P≤0.001; n.s., not significant; Student’s t-test).