Extended Data Fig. 3: Confirmation of TENT4-dependent regulation of PRE reporters.
From: Viral hijacking of the TENT4–ZCCHC14 complex protects viral RNAs via mixed tailing
a, Knockout confirmation by western blotting. GAPDH was used as a loading control. b, RNA levels of firefly reporters harboring subregions of PRE were measured by RT-qPCR (PRE:PREα:PREβ, n = 4; αΔ1, n = 2; αΔ2, n = 1 independent experiment). mRNA levels of both renilla and control vector were used for normalization. Mean was calculated and Error bars represent SEM. *P < 0.05; two-sided t test. c, siRNA knockdown confirmation by western blotting in HEK293T cells. GAPDH was used as a loading control. d, Firefly luciferase activity of PRE reporter constructs in TENT4-depleted HEK293T cells (PRE:PREα:PREβ, n = 7; αΔ1, n = 6; αΔ2, n = 5 independent experiments). Luciferase activities of both renilla and control vector were used for normalization. Mean was calculated and Error bars represent SEM. *P < 0.05, **P < 0.01, ***P < 0.001; two-sided t test. e, Over-expression confirmation by western blotting in KO cells. GAPDH was used as a loading control. f, RNA levels of firefly reporters used in rescue experiments in TENT4 KO cells with ectopically expressed TENT4 wild-type or mutant (n = 3 independent experiments) were measured by RT-qPCR (n = 3 independent experiments). mRNA levels of both renilla and control vector were used for normalization. Mean was calculated and Error bars represent SEM. *P < 0.05, **P < 0.01; two-sided t test. Uncropped blots for panel a,c,e and data for graphs in b,d,f are available as source data.
