Figure 3.

RPL39L KO leads to differentiation defects in E14 mESCs. (A) Representative bright field images showing the spermatogenic differentiation of WT E14 cells. Red arrows indicate spermatocyte-like cells. (B) qRT-PCR assays of DAZL (late) and STELLA (early) sperm cell markers (33) (y-axis, log₂ fold-change) in differentiating (4 days in RA-containing medium) populations of KO clones (x-axis) relative to WT. (C) qRT-PCR of extraembryonic endoderm markers (DAB2, GATA4, GATA6) (65) in RPL39L KO lines relative to WT. (D) Similar for FGF5, NESTIN, and PAX6 ectoderm markers (65). (E) Immunofluorescence staining of embryoid bodies subjected to spontaneous differentiation: GATA4 was used as endoderm marker, NESTIN as ectoderm marker and DAPI to delineate the nucleus. (F) Quantification of GATA4 and NESTIN expression in immunofluorescence images. AFU - arbitrary fluorescence units normalized to DAPI. In all panels, *, ** and *** correspond to P-values <0.05, <0.01 and < 0.001, respectively in the two tailed t-test comparing KO lines with WT.