Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - Stress-induced cytoplasmic accumulation of hnRNP A1 increases FGF-2 protein levels. (A) HeLa cells were grown in high-osmolarity growth medium (0.4 M OSM) or D-MEM for 5 h, nuclear (NE) and cytoplasmic (CE) protein fractions were harvested, separated by SDS-PAGE, transferred to PVDF, and subjected to Western-blot analysis with an antibody against hnRNP A1. (B) Osmotic shock increases FGF-2 protein levels. Cells were treated as described in (A), and total protein extracts were separated by SDS-PAGE, transferred to PVDF, and subjected to Western-blot analysis with antibodies against FGF-2 and actin. (C) Osmotic shock does not affect FGF-2 mRNA levels. HeLa cells were treated with D-MEM (Control) or 0.4 M OSM for 5 h, total RNA was then isolated and cDNA was generated by reverse transcription. Quantitative PCR was used to determine the levels of FGF-2 mRNA and 18S RNA; values are expressed as FGF-2 relative to 18S, and the ratio for control cells was set as 1. Mean +/- S.D. (bars) of three experiments.
• Supplemental Figure 2 - hnRNP A1 does not effectively compete with the La autoantigen for binding to XIAP IRES RNA. GST-hnRNP A1 or GST-La were pre-incubated with ³²P-labeled XIAP IRES RNA probe for 15 minutes and increasing amounts of GST-La or GST-hnRNP A1 were added and incubated for an additional 30 minutes, UV-crosslinked, and then separated by SDS-PAGE and visualized by autoradiography and Coomassie Brilliant Blue staining.