PMC:6288413 / 13969-15219 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/6288413","sourcedb":"PMC","sourceid":"6288413","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6288413","text":"Figure 3 Common Intronic Variant Identified Causes Aberrant Splicing and POLE-Deficient Cells Show Deficiency of Polymerase Epsilon and Slowed S-phase Progression\n(A) The c.1686+32C\u003eG mutation causes aberrant splicing of intron 15 in subject cells. RT-PCR of POLE transcripts from primary fibroblasts. Primers indicated by arrows in schematic. P1, P3, POLE-deficient subjects; C1, C2, control subjects.\n(B) Minigene assay demonstrating that aberrant splicing is a direct consequence of the c.1686+32C\u003eG mutation. +ve control, point mutation in splice donor site, c.1686+1G\u003eA. 5′ \u0026 3′ indicate artificial vector-associated exons.\n(C) POLE1 levels are markedly reduced in subject fibroblasts. Immunoblot of total cell extracts. POLE1 antibody raised against AA1-176. Vinculin, loading control. ∗ non-specific band.\n(D and E) Fibroblast cells from affected individuals exhibit delayed S phase progression. Schematic, experimental set-up.\n(D) Representative FACS plots.\n(E) Quantification of n = 3 affected and n = 3 control cell lines from representative experiment (of n = 3 expts with n ≥ 2 biological replicates per group). Mid-S-phase mean (±SEM) BrdU-labeled cells, normalized to t = 0 time point are plotted for each group. p value, two-way ANOVA.","divisions":[{"label":"label","span":{"begin":0,"end":8}},{"label":"p","span":{"begin":9,"end":162}},{"label":"p","span":{"begin":163,"end":402}},{"label":"p","span":{"begin":403,"end":628}},{"label":"p","span":{"begin":629,"end":812}},{"label":"p","span":{"begin":813,"end":934}},{"label":"p","span":{"begin":935,"end":965}}],"tracks":[]}