Fig. 6: Characterization of rs2790444.
From: Functional dissection of inherited non-coding variation influencing multiple myeloma risk
a Motif analysis predicted that the WAC high-expressing allele rs2790444-T creates a new binding site for POU2F1. Arrow indicates the altered recognition base. b Electromobility shift assay showing selective binding of POU2F1 to rs2790444-T probe. Arrow indicates supershift with an antibody towards POU2F1. c siRNA-knockdown of POU2F1 attenuated luciferase activity for rs2790444-T relative to rs2790444-C in L363 cells. The y-axis indicates the log2 ratio of the luciferase/renilla signal for rs2790444-T relative to the rs2790444-C construct. The bottom, middle, and top of each box plot represent the 25th, 50th, and 75th percentiles. The whiskers represent the non-outlier minimum and maximum values, located at 1.5 times the interquartile range from the bottom and top of the box, respectively. d Western blot confirming knockdown. e Quantitative PCR showing expression of WAC relative to control in MOLP8 cells upon dual-sgRNA CRISPR/Cas9 deletion of a 139-bp region harboring rs2790444, heterozygous for rs2790444. Blue bars indicate the averages of the individual measurements. f Agarose gel confirming deletion of the targeted region. LP labeled probe, NE L363 nuclear extract, ULP unlabeled probe, α-POU2F1 antibody against POU2F1. The P values is for the two-sided Student’s t-test.
