Figure 4.

Validation of translatome data on IFNs and IFN-induced gene expression. Total RNA (Total) and high occupancy (Heavy) polysome-associated mRNAs were extracted from untreated DC (CTRL), DC stimulated with rapamycin for 12 h (RAPA) or DC infected for 16 h with Mtb alone (MTB) or in combination with rapamycin (MTB+RAPA, added 4 h after infection). (A) IFNA1, IFNB1 and IFNL1, (B) CXCL9 and CXCL11, (C) IFIH1, IFIT3 and IRF7 copy numbers were determined by digital PCR. Data are represented as the mean copy number per sample ± standard error of 4 experiments performed with RNAs derived from a set of experiments independent than those used in transcriptome/translatome studies and that yielded similar results. Significance was calculated by analysis of variance (ANOVA) followed by multiple comparison performed with Tukey’s test as specified in Materials and Methods section.