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. 2020 Apr 24;11:1997. doi: 10.1038/s41467-020-15844-2

Fig. 5. XBP1s enhancement of E1A and 19K levels depends on the E1A promoter.

Fig. 5

a The expression of human XBP1s (hXBP1s) enhances E1A and 19K expression and infection. Dose-dependent expression of hXBP1s in HeLa I-KO (IRE1α-knockout) cells transduced with lentivirus vectors for 72 h (first panel). AdV-C2-dE3b-mCherry infection (MOI 100) is rescued in HeLa I-KO cells by hXBP1s expression, where the cells were transduced with lentiviruses for 2 days and infected with AdV-C2 for 24 h (second panel). Data are presented as mean from two technical replicates, and two independent experiments gave similar results. qrt-PCR showing the rescue of E1A mRNA by hXBP1s overexpression in AdV-C2-(MOI 75) infected HeLa I-KO cells. Data are presented as mean ± SD from three independent experiments (third panel). Increased 19K protein levels by hXBP1s expression in AdV-C2-(MOI 75) infected HeLa I-KO cells (n = 3627 and 2317 for lenti-empty and lenti-hXBP1s, respectively, fourth panel). Data show the medians and first and third quartiles, and whiskers as boxes and lines, respectively. Statistical analyses were done by two-tailed Wilcoxon nonparametric tests with significance *p < 0.001. Two independent experiments gave similar results. b E1A expression from the E1A-e/p can be increased by ectopic XBP1s, or reduced by the IRE1α inhibitor 4µ8C in HeLa cells (left panel). A similar experiment was carried out in HDF-TERT cells including IFN-γ to suppress the E1A promoter/enhancer expressed from a lentivirus vector (middle panel). E1A expression in HDF-TERT cells is reduced by RNA interference against IRE1α and XBP1, but not by nontargeting siRNA (siNeg1, 20 nM siPools, right panel). Error bars represent standard deviations. For all the graphs, data are presented as mean from two technical replicates, and two independent experiments gave similar results.