MBNL2 controls the hypoxia response of breast and lung cancer cells by regulating transcript abundance. (A) Western blot of MBNL2 after siRNA-mediated knockdown in hypoxic A549 and MCF-7 cells. Anti-MBNL2 was used to verify the respective knockdown. Total lane protein is shown as loading control. n = 3–4. (B) Venn diagram showing the overlap of differentially expressed genes after knockdown of MBNL2 and in response to hypoxia in MCF-7. P < 1.61 × 10−37 (Fisher's exact test, one-tailed). The number in brackets refers to genes regulated in the opposite direction. (C) Scatter plot comparing the fold changes of shared differentially expressed genes after knockdown of MBNL2 and in response to hypoxia in MCF-7. (D) RT-qPCR quantification of the mRNA levels of HIF target genes in normoxic and hypoxic A549 and MCF-7 cells and after knockdown of MBNL2. Values are normalized to the housekeeping gene RPLP0. n = 3–6. (E) Western blot of MBNL1 and MBNL2 after siRNA-mediated knockdown of MBNL1 in hypoxic MCF-7 cells. Anti-MBNL1 and anti-MBNL2 were used to verify the respective protein levels. Total lane protein is shown as loading control. n = 3. (F) RT-qPCR quantification of the mRNA levels of MBNL1, MBNL2, and HIF target genes. n = 3. (**) P < 0.01. (*) P < 0.05.
