Fig. 2
RP11-367G18.1 variant 2 interacts with YY1 to activate H4K16Ac. (A) RNA pull-down assay and Coomassie blue staining revealed 11 RP11-367G18.1 variant 2-specific bands (left). Protein identity of the 11 bands was analyzed via LC-MS/MS and was shown in a table (right). (B) Knockdown of YY1 suppressed H4K16Ac levels in H1299 cells. (C) Overexpression of YY1 increased H4K16Ac levels in H1299 cells. (D) YY1 was pulled down by biotinylated sense RP11-367G18.1 variant 2. Beads or biotinylated anti-sense RP11-367G18.1 variant 2 were used as the negative control. (E) YY1-induced H4K16Ac activation was suppressed following RP11-367G18.1 variant 2 knockdown. (F) mRNA levels of YY1 were upregulated under hypoxia in FADU and MCF7 cells. (G) Protein levels of YY1 were upregulated under hypoxia in FADU and MCF7 cells. (H) Reporter constructs containing wild-type and mutant HREs in YY1 promoter were shown (left). Reporter assay revealed that HRE (-493/-489) was responsive to hypoxia in YY1 promoter (right). (I) ChIP assay revealed that HIF-1α bound to the YY1 proximal promoters containing HRE under hypoxia. For hypoxic conditions, cells were cultured in 1% O2, 5% CO2, and 94% N2 for 18 h. Scr, Scrambled; Cont, control; V2, variant 2. Data are represented as the mean ± SD. *P < 0.05