Skip to main content
. 2016 Mar 21;113(14):E2047–E2056. doi: 10.1073/pnas.1602883113

Fig. 5.

Fig. 5.

ALKBH5 overexpression promotes BCSC enrichment. (A and B) MDA-MB-231 (A) and MCF-7 (B) subclones transfected with EV or vector encoding ALKBH5 were exposed to 20% or 1% O2 for 48 h, and m6A immunoprecipitation coupled with RT-qPCR was performed to determine the percentage of input NANOG mRNA with m6A methylation (m6A+; mean ± SEM; n = 3; *P < 0.05 and **P < 0.01 vs. EV at 20% O2; #P < 0.05 vs. EV at 1% O2). (C and D) MDA-MB-231 (C) and MCF-7 (D) subclones were exposed to 20% or 1% O2 for 24 h and NANOG mRNA levels were quantified and normalized to EV at 20% O2 (mean ± SEM; n = 3; **P < 0.01 and ***P < 0.001 vs. EV at 20% O2). (E and F) MDA-MB-231 (E) and MCF-7 (F) subclones were exposed to 20% or 1% O2 for 48 h, and immunoblot assays were performed. NANOG protein expression was quantified and normalized to EV at 20% O2 (mean ± SEM; n = 3; **P < 0.01 vs. EV at 20% O2). (G and H) MDA-MB-231 (G) and MCF-7 (H) subclones were exposed to 20% or 1% O2 for 72 h, and the percentage of ALDH+ cells was determined (mean ± SEM; n = 3; **P < 0.01 and ***P < 0.001 vs. EV at 20% O2; #P < 0.05 vs. EV at 1% O2).