Figure 4 | Scientific Reports

Figure 4

From: Differential antitumor activity of compounds targeting the ubiquitin-proteasome machinery in gastrointestinal stromal tumor (GIST) cells

Figure 4

Delanzomib (DLZ) treatment leads to upregulation of soluble histone H2AX but downregulates KIT expression through inhibition of transcription. (A,B) Immunoblot analysis of GIST cells treated with delanzomib at the indicated concentrations for 72 h (A) or with 0.1 μM delanzomib for the indicated times (B) and probed for phospho-H2AX (S139) and total H2AX as well as phospho-KIT (Y719) and total KIT. Grouped immunoblot images are either cropped from different parts of the same gel or from a separate gel run with another aliquot of the same protein lysate. (C,D) RT-PCR (C) and quantitative RT-PCR (qRT-PCR) amplification (D) of KIT mRNA after treating GIST cells with DMSO or 0.1 μM delanzomib in comparison to bortezomib (BO) for 48 h. GIST882 cells were also treated with the RNA polymerase II inhibitor α-amanitin (α -ama) or H2O solvent control. (E) Immunofluorescence microscopic analysis of GIST882 and GIST48 cells treated with DMSO or 0.1 μM delanzomib for 72 h and stained for the transcriptional co-activator CREB-binding protein (CBP; green). Nuclei were stained with DAPI. Bar, 20 μm.

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