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. 2016 Oct 26;7(48):78281–78296. doi: 10.18632/oncotarget.12930

Figure 4. AZD induces HSP70 protein expression without interfering with the functions of HSP90 or proteasome.

Figure 4

WCE (20 μg) of HEK293 cells pretreated for 16 h with bortezomib (BTZ) or geldanamycin (Gel) or various concentrations of AZD, or DMSO were subjected to immunoblot A. using the antibodies against anti-ubiquitin (P-Ub). The levels of HSP70 protein in the same samples were also determined as indicated using anti-HSP70 antibody. B. The bands in the blot were estimated and plotted as bar graph considering ß-actin levels as an internal loading control. C. Immunoblot with antibodies against two HSP90 client proteins Akt or Raf1 and HSP70 and HSP90 itself to compare their relative expression levels in those samples. D. Band intensities in the blot were estimated by densitometric scanning and plotted to compare their expression levels. ß-actin levels were used as internal loading control.