(A) Scheme depicting the dsRNA induced innate immune response pathways.
(B) Q-RT-PCR determination of expression of dsRNA sensor genes Ifih1 (MDA5) and Ddx58 (RIG-I) in E14.5 fetal livers, normalized to Gapdh (n=3 biological replicates per group).
(C-D) Detection of PKR phosphorylation and eIF2α phosphorylation in fetal liver in response to Mettl3 deletion via Western blot (C); bands from three representative blots were quantified via Image J, p-PKR and p-eIF2a protein expressions were normalized to PKR and eIF2a expression, respectively (D) (n=3 biological replicates per group).
(E) RtcB Q-RT-PCR analysis of tRNA specific cleavage by RNase L, normalized to U6 (n=3 biological replicates per group).
(F) Q-RT-PCR determination of expression of immune response genes downstream of interferon activation in E14.5 fetal livers, normalized to Gapdh (n=3 biological replicates per group).
(G) Q-RT-PCR determination of IFN III (Ifnl3) and I (Ifna4, Ifnb1) expression in E14.5 fetal livers, normalized to Gapdh (n=3 biological replicates per group).
Data are represented as mean ± SEM, and representative of at least three independent experiments; The P values were calculated using two-tailed Student's t test. * p<0.05, ** p<0.01.
See also Figure S6.