Hypoxia causes CDH22 protein accumulation in an eIF4E2-dependent manner independent of its transcript abundance. (a–d) Western blot of CDH22 and eIF4E2 protein levels in normoxic (21% O2) and hypoxic (1% O2) U87MG (a, c) and MDA-MB-231 (b, d) control cells stably expressing a non-targeting shRNA, in cells stably expressing shRNA targeting eIF4E2 mRNA (KD1.1 or 1.2), KD cells stably expressing an exogenous empty vector (KD Ctrl) or the eIF4E2 coding sequence (Exo1 4E2). GAPDH used as a loading control. (e, f) CDH1 (e) and CDH22 (f) mRNA levels measured by quantitative reverse transcriptase–PCR in U87MG and MDA-MB-231 control cells stably expressing a non-targeting shRNA exposed to normoxia or hypoxia. Data are presented as mean ±s.e.m., n =3, *P<0.05 (Student’s t-test). (g) Immunofluorescence of CDH22 in U87MG and MDA-MB-231 cells exposed to normoxia and hypoxia. Scale bar, 10 μm.