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. 2016 Jul 27;6:30406. doi: 10.1038/srep30406

Figure 5. HDAC6 is implicated in regulating exosome-dependent GRP78 secretion.

Figure 5

(A) Relative mRNA levels of HDAC1 and HDAC6 in normal colon FHC cells and colon cancer cells (HCT-116, DLD1, SW480, HT-29 and SW620). (B) Western blot analysis of HDAC6 in normal mouse intestine tissue or intestinal tumour tissue from an AOM/DSS-induced colon tumourigenesis model. (C) DLD1 cells stably expressing GRP78-GFP were treated with tubastatin A and immunostained with an anti-CD63 antibody. The superimposed confocal images show the colocalization of GRP78 and CD63. (D) DLD1 cells stably expressing GFP and GRP78-GFP were treated with tubastatin A and immunostained with an anti-HDAC6 antibody to analyse the relative intracellular location alterations of GRP78 and HDAC6. (F) HDAC6 expression in DLD1 cells was knocked down by shRNAs. Culture supernatants were collected and subjected to Western blot analysis to assess GRP78 secretion. Western blot analysis of HDAC6 expression was performed to confirm the knockdown efficiency of the shRNAs.