Fig. 2.

Hypoxia affects LCMV transcription and protein levels. (A) RLM-RACE analysis was performed on RNA isolated from hypoxic and normoxic HeLa-MX cells. Capped RNA transcripts were determined by semiquantitative RT-PCR using a β-actin internal standard. Data show fold induction of viral mRNA levels in hypoxia as in Fig. 1. (B, left) Immunoblot analysis of the indicated viral proteins with specific antibodies using whole-cell extracts prepared from HeLa-MX cells cultured for 48 h under normoxia (NO) or hypoxia (HY). The signal obtained with an antiactin antibody was used as a control for loading and transfer efficiency. Detection of HIF-1α served as a control for the induction of a cellular response to hypoxia. One representative of at least three independent experiments with similar results is shown. (B, right) The intensity of protein bands was evaluated with ImageJ 1.34s software. The relative quantity of proteins was calculated as the ratio of the intensity of each band to the intensity of the related actin internal standard.