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. 2021 Oct 25;22(21):11505. doi: 10.3390/ijms222111505

Figure 2.

Figure 2

NCAPH was one of the OCT1-regulated genes in MCF-7 cells. (A) The results of microarray analyses were summarized. Genes downregulated with a fold change ≥8 by silencing OCT1 with siOCT1 #1 (blue oval) or with siOCT1 #2 (red oval) compared to treatment with siControl are shown. The digits indicate the number of genes. Sixteen genes were downregulated in common. (B) Western blot analysis for NCAPH and OCT1 expressions in MCF-7 cells and LTED cells treated with two kinds of siRNAs for OCT1 (siOCT1 #1 or #2) or siControl (siCont.). β-actin protein was blotted as a loading control. IB, immunoblot. (C) Western blot analysis for NCAPH and OCT1 expressions in two clones of MCF-7 cells stably expressing OCT1 (OCT1-OE #1 and #2) and an MCF-7 clone transfected with empty vector (Vector). β-actin protein was blotted as a loading control. (D) Schema of NCAPH promoter region. A putative octamer consensus sequence (ATTTAAAA) exists at 26 base pairs upstream from the translation initiation site (ATG) of NCAPH gene. (E) Association of OCT1 in the promoter region of NCAPH in MCF-7 cells. ChIP assay for OCT1 or normal rabbit IgG was performed. The fold enrichments relative to IgG in NCAPH promoter and another locus in NCAPH (negative control) were measured by performing quantitative PCR (qPCR). Relative fold enrichment was shown as mean and SEM (n = 3). * p < 0.05.