Journal: The FASEB Journal

Article Title: MS 275 Inhibits Neuroblastoma Cell Growth by Mediating H 3 K 27ac/ PROX 1 Axis In Silico and In Vitro

doi: 10.1096/fj.202500464RR

Figure Lengend Snippet: MS275 increases PROX1 expression by inducing H3K27 acetylation. (A) The H3K27ac enriched area at the promoter of PROX1 was analyzed online at the UCSC Genome Browser ( http://genome.ucsc.edu ). (B, C) ChIP assays. The SH‐SY5Y cells were cultured in a 100‐mm dish and treated with 0 or 10 μM MS275 for 48 h. Then the cells were subjected to ChIP assay with anti‐H3K27ac antibody. (B) The interaction between H3K27ac and PROX1 promoter was detected and quantified using qPCR analysis. (C) The qPCR products were analyzed with agarose gel electrophoresis. (D) SH‐SY5Y cells were similarly treated as in Figure , and cells were harvested for western blot analysis with H3K27ac antibody. Compared to 0, each experiment was repeated at least three times independently. * p < 0.05; ** p < 0.01. (E) SH‐SY5Y cells were cultured in 6 wells and transfected with three interference RNAs of HDAC1 (flag as HDAC1#1, HDAC1#2, and HDAC1#3), three interference RNAs of HDAC3 (flag as HDAC3#1, HDAC3#2, and HDAC3#3), and their control siNC. Forty‐eight hours later, cells were collected for western blot analysis with PROX1 antibody. Each experiment was repeated at least three times independently. * p < 0.05;*** p < 0.001; **** p < 0.0001; NS, no significant. (F) The interference efficiency of HDAC1 and HDAC3 siRNAs. SH‐SY5Y cells were treated as in E and verified the interference efficiency of HDAC1 and HDAC3 using Western blot with indicated antibodies.

Article Snippet: The monoclonal antibodies (mAbs) against HDAC1, HDAC3, Phospho‐CDK2 (Tyr15), CDK2, Phospho‐RB (Ser807), and CDK4 were from MedChemExpress.

Techniques: Expressing, Cell Culture, Agarose Gel Electrophoresis, Western Blot, Transfection, Control