Hypoxia influences polysome distribution of human ribosomal protein S12 and alternative splicing of ribosomal protein mRNAs

Endogenous RPS24 short and long transcript variants, and exogenous protein isoforms associate with polysomes. (A) Using transcript variant-specific RPS24 primers, short and long variants were amplified via RT-PCR from polysome fractions with either cycloheximide, or EDTA as a control to dissociate polysomes. (B) Polysomes were isolated from two cell lines each stably expressing one of the FLAG-tagged RPS24 protein isoforms (eS24) using cycloheximide and EDTA. Western blots were performed using anti-FLAG, or β-Actin as a control protein not associated with polysomes. Experiments performed in U87MG glioblastoma in normoxia (21% O₂). See Supplemental Table S1 for primer sequences.