Fig. 2

LINC00115 activates the HIF1α pathway and thereby enhances BCSC properties. A A volcano plot of differentially expressed genes in LINC00115 KD MDA-MB-231 BCSCs (derived from MDA-MB-231/PTX_R cells) with LINC00115 shRNA-2 (sh115-2). The red dots indicate the significantly upregulated genes and the blue dots indicate the significantly downregulated genes. B Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of downregulated pathways by LINC00115 KD. C GSEA analyses show that LINC00115 KD downregulates the HIF1 signaling pathway. NES, normalized enrichment score. D Western blotting (WB) of effects of LINC00115 KD on HIF1 signaling inactivation in MDA-MB-231 and BT549 BCSCs. E QRT-PCR of expression of HIF1α, LDHA, and MDR1. F Ectopic expression of HIF1α rescues mammosphere formation inhibited by LINC00115 KD. G Limiting dilution mammosphere-forming assays were determined and calculated in cells from panel (D). H Cell viability analysis. BCSCs derived from MDA-MB-231/PTX_R and BT549/PTX_R cells from D were treated with paclitaxel for 72 h. Cell viability was determined with Sulforhodamine B colorimetric (SRB) assay. I Cells generated in D were subjected to transwell assays and relative invasion numbers were calculated. J QRT-PCR of effects of LINC00115 depletion on LINC00115 levels in metastatic lung tissues from Fig. 1G. K WB of effects of LINC00115 depletion on HIF1α, LDHA, and MDR1 levels in metastatic lung tissues from Fig. 1G. Data are representative of three independent experiments with similar results. Error bars, ± SEM. **P < 0.01, and ***P < 0.001, by paired two-tailed t-test