γ secretase inhibitor dapt  (Cell Signaling Technology Inc)

Journal: Science Advances

Article Title: HDAC7 promotes renal cancer progression by reprogramming branched-chain amino acid metabolism

doi: 10.1126/sciadv.adt3552

Figure Lengend Snippet: ( A ) KEGG pathway analysis showing pathways positively correlated with SNAI1 expression in metastatic RCC samples ( n = 26). MAPK, mitogen-activated protein kinase; cAMP, cyclic adenosine 3′,5′-monophosphate; cGMP, guanosine 3′,5′-monophosphate; PKG, cGMP-dependent protein kinase. ( B ) Relative mRNA expression of SNAI1 in 769-P cells treated with indicated inhibitors for 48 hours. ( C ) Relative mRNA expression of SNAI1 in 769-P cells incubated with or without 25 μM DAPT. DAPT-treated cells were exposed to media with or without BCAA for 16 hours ( n = 3). ( D ) Immunoblot analysis of SNAIL1 expression in 769-P cells treated with specific inhibitors, followed by exposure to media with or without BCAA for 16 hours. ( E and F ) Western blot analysis of indicated proteins in RXF-393 cells (E) and 786-O cells (F) treated with increasing concentrations of BCAA for 16 hours. ( G ) Western blot analysis of indicated proteins in 786-O cells expressing shNC or two independent shRNA targeting HDAC7 . ( H ) Western blot analysis of indicated proteins in 786-O cells expressing shNC or two independent shRNA targeting HDAC7, treated with increasing concentrations of BCAA for 16 hours. In (B) and (C), data are presented as means ± SEM and are representative of at least two independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test was used for (B) and (C). ** P < 0.01 and *** P < 0.001.

Article Snippet: The γ-secretase inhibitor DAPT was obtained from MilliporeSigma.

Techniques: Expressing, Incubation, Western Blot, shRNA