Fig. 6: Flow cytometry and qRT-PCR validation of eIF5a-regulated proteins. | Nature Communications

Fig. 6: Flow cytometry and qRT-PCR validation of eIF5a-regulated proteins.

From: Translation factor eIF5a is essential for IFNγ production and cell cycle regulation in primary CD8+ T lymphocytes

Fig. 6

a Intracellular stains for eIF5a or hypusine were used to identify WT (blue line) and KO (red line) populations in Eif5a, Dhps, or Dohh CRISPR-KO cells (dot plots on left show the extent of KO). Thy1 CRISPR cells (grey shaded line) were used as a KO control. Overlay histograms on right show IRF4, TBET, EOMES and CDK1 expression in electronically gated WT and KO samples. b Quantification of multiple paired samples from the analysis in (a) (P values were calculated using paired two-sided t test). Dots: blue, WT; red, KO. c mRNA of these genes were quantified by RT-qPCR, normalised by the level of Actb gene, in fresh bulk transfected cells (n = 3 biological samples, P values were calculated using unpaired two-sided t test). Centre line and error bars represent mean value ±SD between biological replicates, respectively. P value colours: blue, significant; black, non-significant.

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