Figure 2.

Hsp90 was up-regulated and affected its client proteins in heat-stressed CMVECs. Western blot analysis and immunocytochemistry were performed with the indicated antibodies. Data represent the means ± SD for three independent experiments. (A) CMVECs were treated with or without heat stress of the designed times. Total proteins were used for semi-quantitative detection of the corresponding proteins. The relative abundance of the tested proteins was normalized to that of β-actin. (B) CMVECs were treated with or without heat stress of the designed times. Immunofluorescence was performed using anti-Hsp90α, anti-Akt, anti-PKM2, and anti-Hsp70 antibodies. Representative images are presented (Hsp90: red fluorescence, Akt/PKM2/Hsp70: green fluorescence, nucleus: blue fluorescence, the merged signals of Hsp90 with Akt/PKM2/Hsp70 in the cytoplasm: yellow fluorescence). Bar = 20 µm. The differences of the data of cells with different heat stress times vs. that of the non-stressed cells are indicated by * p < 0.05 and ** p < 0.01.