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. 2018 Nov 14;8:16820. doi: 10.1038/s41598-018-34887-6

Figure 1.

Figure 1

P140 affects the nuclear translocation of HSPA8 upon heat shock. (A) Confocal fluorescence images of cellular HSPA8 upon HS and recovery in the presence or absence of P140. MRL/N-1 cells were exposed to HS (30 min; 45 °C), and allowed to recover at 37 °C for indicated time points (3 and 24 h) following HS, in the absence or presence of 10 µM P140. Cells were stained with HSPA8-PE antibody (in red). The nuclei were stained with DAPI (in blue). Transmission images showing the morphology of cells are presented in Fig. S1A. (B) Western immunoblotting of nuclear HSPA8 upon HS and recovery in the presence or absence of P140. MRL/N-1 cells were treated and allowed to recover as in (A). Cells were collected at the indicated time points of the recovery phase and subjected to cell fractionation to isolate the nuclear and cytoplasmic fractions. Nuclear HSPA8 only was analyzed here. Histone H3 was used as nuclear marker and β-tubulin as cytosolic marker. The relative amount of nuclear HSPA8 was analyzed by densitometry with Image J. The fold change was plotted by normalizing the nuclear HSPA8 amounts by that of the control after HS (value set to 1). Mean values with standard error of the mean (SEM) of five independent experiments are presented. Non-parametric two-way ANOVA was used to evaluate the statistical significance.