Fig. 4

p65 positively regulates ethanol-induced STARD10 and ERBB2 expression binding their promoter sequences in MCF-7 cells. a STARD10 and ERBB2 promoter activity assay and RT-PCR for STARD10 and ERBB2 mRNA levels were measured in MCF-7 cells treated with 100 mM ethanol and transfected with sip65 (10 nM) for 48 h.. *p < 0.03 vs. Sc. †p < 0.04 vs. EtOH for STARD10 promoter; *p < 0.04 vs. Sc. †p < 0.04 vs. EtOH for ERBB2 promoter. *p < 0.004 vs. Sc; †P < 0.03 vs. EtOH for mRNA levels. b mRNA levels of STARD10, ERBB2 and RELA measure by RT-PCR from 3 indenpendet experiments. *p < 0.004 vs. Sc; †p < 0.03 vs. EtOH. c Western blotting analysis of p65 in nuclear and cytoplasmic fractions. Nuclear marker (H3) and cytosolic marker (tubulin) were immunoblotted to demonstrate fraction purity. Data are expressed as (mean ± SE) from triplicate of 4 independent experiments. *p < 0.05 vs. control. d ChIP analysis for p65 binding to STARD10 and ERBB2 promoters. Input genomic DNA (Input DNA) was used as a positive control, while non-specific antibody IgG was used as a negative control. Results are summarized as the densitometric changes as fold of control after normalizing to input DNA. *p < 0.03 vs. control. e Western blotting analysis of p65 from nuclear and cytoplasmic fractions from ethanol-fed MMTV-neu transgenic mice tumor. Results are expressed as fold relative to control tumor (mean ± SE) from 4 mice per group. *p < 0.05 vs. control tumor. f ChIP analysis of p65 binding to ERBB2 and STARD10 promoters in MMTV-neu ethanol-fed transgenic mice tumor. Results are summarized as the densitometric changes as fold of control after normalizing to input DNA. *p < 0.04 vs. control