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Fig. 1 | BMC Cancer

Fig. 1

From: Generation of a PAX6 knockout glioblastoma cell line with changes in cell cycle distribution and sensitivity to oxidative stress

Fig. 1

PAX6 is knocked out by CRISPR-Cas9 technology in U251 N glioblastoma cells and reintroduced by an EGFP-PAX6 expressing plasmid (a) Western blot on cell lysates originating from single cell clones after CRISPR-Cas9 treatment. PAX6 Control cells have undergone CRISPR-Cas9 treatment and single cell sorting but still express PAX6. Detection by anti-PAX6 antibody. Actin staining was used for loading control. KO, knock out; WT, wild type U251 N; L, Molecular weight marker. b Immunocytochemistry (ICC) on WT U251 N and pooled PAX6 KO cell lines (2A.3, 2A.28 and 2.10). Anti-PAX6 antibody detect endogenous PAX6 protein, DAPI staining identifies the nucleus. Scale bars indicate 10 μm. c Western blot on the individual PAX6 KO cell lines transduced with a Dox inducible EGFP-PAX6 expressing plasmid (2A.3R, 2A.28R and 2.10R). The right panel show cells treated with 10 ng/ml Dox for 24 h. Anti-PAX6 and anti-Actin antibodies were used for detection. d ICC of Dox treated pooled Rescue cells expressing EGFP-PAX6. EGFP is detected as green fluorescence, anti-PAX6 antibody stain PAX6 protein, and the nucleus is stained by DAPI in blue. Scale bar indicates 20 μm. Western blots and ICCs were performed on more than three samples of each cell line and showed the same results

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