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. Author manuscript; available in PMC: 2024 Sep 15.
Published in final edited form as: Clin Cancer Res. 2024 Mar 15;30(6):1175–1188. doi: 10.1158/1078-0432.CCR-23-2368

Figure 5. Inhibitory effects of NTX-301 on SCD in OC cells.

Figure 5.

(A) UCSC Genome Browser on Human GRCh38/hg38 tracks of H3K27Ac peaks in the SCD promoter in OVCAR5 cells (33) and other cell lines previously recorded in the ENCODE dataset (GM12878, and HEK293T) (34, 35). The H3K27Ac binding motif along with the position of primer sequences (Primer_2) used for q-PCR are shown. Amplification of a sequence 1 kb downstream was used as a control (Primer_1). (B) ChIP-PCR shows H3K27Ac enrichment in the SCD gene (Primer_2) in OVCAR5 cells treated with DMSO or NTX-301 (100nM, 2days, n=3). Enrichment of H3K27Ac on a sequence 1 kb downstream was used as a control (Primer_1). Results are means ± SD, which was analyzed by two-way ANOVA with multiple comparisons. For all comparisons, *p<0.05; **p<0.01; ***p<0.001. (C) Western blot of H3K27Ac and H3 in OVCAR5 OC cells treated with DMSO or NTX-301 for 2 days. (D, E) mRNA expression levels (mean fold-change ± SD) of SIRT1 measured by qRT-PCR (D) and western blot of SIRT1 and beta actin (loading control) (E) in OVCAR5 cells treated with DMSO or NTX-301 for 2 days (n=3 replicates). (F) DNA methylation levels for 3 significantly demethylated CpG sites (p-adj< 0.05) in the SIRT1 gene as measured with an Illumina EPIC array in OVCAR5 cells treated with DMSO or NTX-301 (100nM, 3 days; n = 3 replicates per group).