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. 2024 Jan 18;22(4):373–385. doi: 10.1158/1541-7786.MCR-23-0629

Figure 5.

Figure 5. Skp2 is a target of AIbZIP. A, Western blot analysis for SKP2, AKT and its phosphorylated form (S473), SRC and its phosphorylated form (Y419) in MFM223 and MDAMB453 cells transfected with Aibzip siRNA. B, Western blot analysis of p27 and SKP2 in MFM223 and MDAMB453 cells under non-treatment and AIbZIP-introduced conditions. C, Quantitative PCR analysis for Skp2 mRNA in MFM223 and MDAMB453 cells transfected with Aibzip siRNA. mRNA levels were normalized by Actb (mean ± SD, n = 4). D, Schematic of the Skp2 promoter region and reporter constructs. Numbers indicate the distance from the transcriptional start site. Luc, luciferase gene, TSS, transcriptional start site. E, Luciferase assays of MFM223 and MDAMB453 cells introduced AIbZIP or transfected with Aibzip siRNA (mean ± SD, n = 4). F, Schematic of the Skp2 promoter and the annealing sites of the primer set used in the chromatin immunoprecipitation assay. Numbers indicate the distance from the transcriptional start site. G, The results of PCR amplification of the Skp2 promoter containing the CRE-binding site after immunoprecipitation by the indicated antibodies. Mouse IgG and Histone H3 were used as negative and positive control, respectively.

Skp2 is a target of AIbZIP. A, Western blot analysis for SKP2, AKT and its phosphorylated form (S473), SRC and its phosphorylated form (Y419) in MFM223 and MDAMB453 cells transfected with Aibzip siRNA. B, Western blot analysis of p27 and SKP2 in MFM223 and MDAMB453 cells under non-treatment and AIbZIP-introduced conditions. C, Quantitative PCR analysis for Skp2 mRNA in MFM223 and MDAMB453 cells transfected with Aibzip siRNA. mRNA levels were normalized by Actb (mean ± SD, n = 4). D, Schematic of the Skp2 promoter region and reporter constructs. Numbers indicate the distance from the transcriptional start site. Luc, luciferase gene, TSS, transcriptional start site. E, Luciferase assays of MFM223 and MDAMB453 cells introduced AIbZIP or transfected with Aibzip siRNA (mean ± SD, n = 4). F, Schematic of the Skp2 promoter and the annealing sites of the primer set used in the chromatin immunoprecipitation assay. Numbers indicate the distance from the transcriptional start site. G, The results of PCR amplification of the Skp2 promoter containing the CRE-binding site after immunoprecipitation by the indicated antibodies. Mouse IgG and Histone H3 were used as negative and positive control, respectively.