ASXL1 Antibody (6E2)

Western Blot: ASXL1 Antibody (6E2) [H00171023-M05] -

Western Blot: ASXL1 Antibody (6E2) [H00171023-M05] - Functional effects of CRISPR/Cas9-mediated ASXL1 mutation correction(A) Evaluation of ASXL1 protein expression by WB. Left: ASXL1 protein expression in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, the K562 leukemia cell line (carrying the Y591X heterozygous ASXL1 mutation), & KBM5 clones (labeled 1–5) w/ heterozygous precise correction of ASXL1 mutation. Right: ASXL1 protein expression in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, & KBM5 clones (labeled 1–5) w/ homozygous precise correction of ASXL1 mutation. beta-actin used as loading control. (B) Evaluation of expression levels of HOXA genes using quantitative real-time PCR (q-RT-PCR). Expression levels of HOXA5, 6, 7, 9, 10 & 13 measured in three ASXL1 homozygous corrected KBM5 clones compared w/ uncorrected cells. Results shown obtained from six independent experiments for each clone. Values in ASXL1 homozygous corrected cells are relative to uncorrected cells. Bar graphs show mean + standard error of mean (s.e.m.) (* = P < 0.05, ** = P < 0.01, *** = P < 0.001, paired t-test). (C) Evaluation of H3K27me3 levels & expression of PRC2 components by WB in uncorrected KBM5 cells & three KBM5 clones w/ homozygous correction of ASXL1 mutation. H3K27me3 levels & total H3 levels evaluated using purified histone fractions. Expression levels of two PRC2 components (EZH2, SUZ12) determined using whole cell lysates. beta-actin used as loading control. (D) Immunoprecipitation of BAP1 in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, & three KBM5 clones w/ homozygous correction of ASXL1 mutation. The BAP1 protein fraction immunoprecipitated using a BAP1 antibody & stained for ASXL1 & BAP1. Image collected & cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.6392), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ASXL1 Antibody (6E2) [H00171023-M05] -

Western Blot: ASXL1 Antibody (6E2) [H00171023-M05] - Functional effects of CRISPR/Cas9-mediated ASXL1 mutation correction(A) Evaluation of ASXL1 protein expression by WB. Left: ASXL1 protein expression in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, the K562 leukemia cell line (carrying the Y591X heterozygous ASXL1 mutation), & KBM5 clones (labeled 1–5) w/ heterozygous precise correction of ASXL1 mutation. Right: ASXL1 protein expression in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, & KBM5 clones (labeled 1–5) w/ homozygous precise correction of ASXL1 mutation. beta-actin used as loading control. (B) Evaluation of expression levels of HOXA genes using quantitative real-time PCR (q-RT-PCR). Expression levels of HOXA5, 6, 7, 9, 10 & 13 measured in three ASXL1 homozygous corrected KBM5 clones compared w/ uncorrected cells. Results shown obtained from six independent experiments for each clone. Values in ASXL1 homozygous corrected cells are relative to uncorrected cells. Bar graphs show mean + standard error of mean (s.e.m.) (* = P < 0.05, ** = P < 0.01, *** = P < 0.001, paired t-test). (C) Evaluation of H3K27me3 levels & expression of PRC2 components by WB in uncorrected KBM5 cells & three KBM5 clones w/ homozygous correction of ASXL1 mutation. H3K27me3 levels & total H3 levels evaluated using purified histone fractions. Expression levels of two PRC2 components (EZH2, SUZ12) determined using whole cell lysates. beta-actin used as loading control. (D) Immunoprecipitation of BAP1 in SET2 leukemia cell line (wild-type for ASXL1), uncorrected KBM5 cells, & three KBM5 clones w/ homozygous correction of ASXL1 mutation. The BAP1 protein fraction immunoprecipitated using a BAP1 antibody & stained for ASXL1 & BAP1. Image collected & cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.6392), licensed under a CC-BY license. Not internally tested by Novus Biologicals.