Effect of SLFN11 phosphorylation site mutations on its translation-inhibitory properties.
a, V5-tagged SLFN11 was expressed in HEK293T cells and immunoprecipitated. The indicated post-translational modifications were identified by mass spectrometric analysis. b, the indicated SLFN11 mutants were expressed in HEK293T, and p24 protein derived from cotransfected pNL4–3.Luc.R+E− was determined by anti-p24 Western blotting. c, HIV p24 protein levels from b were quantified and normalized over HIV p24 mRNA (biological replicates, mean ± S.D. (error bars), n = 3). d, HEK293T cells were cotransfected with plasmids encoding the indicated SLFN11 mutants, pNL4–3.Luc.R+E−, V5-tagged wtGFP, and Myc-tagged EGFP, and resulting protein levels were visualized by Western blotting. e, wtGFP and EGFP mRNA levels of d quantitated by qPCR (biological replicates, mean ± S.D. (error bars), n = 3). RQ, relative quantification.