Extended Data Fig. 9: PRC1 is a BL host factor important for silencing of LMP1 and LMP2a expression.
From: DNA methylation enzymes and PRC1 restrict B-cell Epstein–Barr virus oncoprotein expression
a, Volcano plots showing the -Log10 (p-value) statistical significance (y-axis) and Log2 fold-change (x-axis) calculated by STARS analysis of sgRNA abundance in the input versus FACSorted ICAM-1high/CD10low population. Values for genes encoding the PRC1 subunits SCML2 and RING1 are highlighted in blue, and values for genes encoding UHRF1 and DNMT1 are shown in purple for comparison. n = 3 of biologically independent replicates. P values were determined by one-sided Fisher’s exact test. b, Mean +/- SEM normalized SCML2 mRNA reads from triplicate RNAseq analysis of MUTU I versus III (left) or of MUTU I that expressed control or UHRF1 sgRNA (right). n = 3 of biologically independent replicates. P values were calculated by unpaired two-sided student’s t-test with equal variance assumption. c, Immunoblot analysis of WCL from MUTU III or from MUTU1 that expressed the indicated sgRNAs. d, qPCR analysis of EBNA3C expression. Values were normalized by 18s rRNA qPCR levels and the ratio in control cells was set to 1. Mean ± SEM from n = 3 biologically independent replicates are shown. p-values were calculated using one-way ANOVA with Sidak’s multiple comparisons. e, Immunoblot analysis of WCL from MUTU III or Daudi that expressed the indicated sgRNAs. f, Fold change of the LMP1:GAPDH immunoblot ratio (left) or BCL6:GAPDH immunoblot ratio (right) from n = 3 of biologically independent replicates, as in ED Fig. 9e. p-values were calculated using one-way ANOVA with Sidak’s multiple comparisons. g, Immunoblot analysis of WCL from MUTU III or from REH acute lymphoblastic leukemia EBV-negative B-cells or from Akata EBV-negative BL expressing the indicated sgRNAs. h, Immunoblot analysis of WCL from MUTU I that expressed the indicated sgRNAs. i, Normalized IRF4 mRNA reads from n = 3 of biologically independent triplicate RNAseq datasets of MUTU I cells that expressed sgControl or sgUHRF1. p-values were calculated by unpaired two-sided student’s t-test with equal variance assumption. Blots in c, e, g and h are representative of at least 3 biologically independent replicates.
