Changes in cellular level of polypeptides during heat shock and recovery.

<p>A) Cells were grown to 40% confluence and heat shocked for 3 hours at 44°C and recovered at 37°C for 24 hours. The control cells were maintained at 37°C. For all experiments cells were directly lysed on the plate using gel loading buffer, and lysates were analyzed by SDS/PAGE. Individual polypeptides were detected by western blotting using appropriate antibodies. B) Shows dose response of western blotting of cell extract from exponentially growing cells with different antibodies. The sample volumes are indicated at the top of the panel. The abundance of specific polypeptides in both panels was determined by scanning the X-ray film and quantifying the images with Image J as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064171#s2" target="_blank">materials and methods</a>. The values in arbitrary scale are shown bellow each lane. All values were normalized by the using the β -actin levels as loading controls. The experiment was repeated three times, and the averages are shown in panel B as mean ± standard error. C) Western blotting and quantification was performed to measure the abundance of PABP4 in control, heat- shocked and recovered cells as described above. Data are mean ± SE, p<0.05 for those with an * and p>0.05 for those that are unmarked, n = 3.