Figure 1. Figure1: HIF-1α and miR-210 expression by myeloma cell lines under hypoxic conditions and validation of permanent HIF-1α knockdown by shRNA. A) Relative expression of HIF-1α mRNA at different hours of hypoxia (1 % and 5 % O2) was evaluated in U266 and RPMI-8226. B) miR-210 expression at different hours of hypoxia (1 %O2) was studied in U266 and RPMI-8226. C, D) Anti-HIF-1α Lentivirus shRNA pool was used for HIF-1α stable knockdown in RPMI8226, whereas the pLKO.1 lentiviral vector was used as the empty control vector. HIF1α mRNA expression was evaluated by real-time quantitative PCR (C) and qualitative PCR (D). E) Value of miR-210 was elevated in RPMI-8226- anti-HIF-1α cells under hypoxic conditions (1 % O2) and RPMI-8226-pLKO.1 (control) at 24, 48 and 72 h. Internal control for all experiments was Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. Columns, mean (n=3); bars, SEM. *P<0.05, **P<0.01, ***P<0.01. Abbreviations: h (hours).
