Supplementary Figure 2: Identification of hypoxia responsive miRNAs associated with tumor progression.

(a) Heat map of small RNA sequencing results from MCF7 cells treated with normoxia or hypoxia, MCF7 cells overexpressing HIF-1α, and MCF7-derived BTIC cells. (b) The correlation of selected hypoxia responsive miRNAs with breast tumor progression. Red, molecule is up-regulated. Green, molecule is down-regulated. The intensity of green and red molecule color indicates the degree of down-regulation and up-regulation, respectively. Font bold in red indicates the focus molecule. (c) The correlation of selected hypoxia responsive miRNAs with certain key cancer signaling pathways. (d) The expression level of miR106b-25 cluster in normoxic, hypoxic, HIF-1α overexpressing, and BTIC cells. MCF7 and MDA-MB-231 cells were used as models, as indicated. (e) Knockdown of HIF-1α abolished hypoxia effect on induction of miR25/93 in either hypoxic MCF7 (left upper) or MDA-MB-231 cells (left lower). The protein levels for HIF-1α were presented in right panel. The cells expressing shRNA targeting EGFP were used as control. All the data in the graphs are presented as means ± SD (n = 3 independent experiments). Samples were compared using two-tailed Student’s t test. Asterisk indicates P < 0.05, compared with controls. The immunoblotting shown in panel e are representative of three independent experiments. Unprocessed scans of western blot analysis is available in Supplementary Figure 9. Statistics source data is available in Supplementary Table 4.