Fig. 6: Pazopanib and AZD5153 exhibits pro-apoptotic effects by supressing oncogenic MYC pathways.

a Principal component analysis (PCA) of log₂ fold change in gene expression of untreated and treated groups of LPS141 and hSC40 across three biological replicates. b GSEA analysis to identify significantly enriched hallmark pathways. Pathways with a fold change (FC) of ≥1 and P < 0.05 were considered significantly altered. AvsU, AZD5153 vs Untreated; PvsU, Pazopanib vs Untreated; CvsU, Combined vs Untreated. c Heatmap of log₂ fold change in gene expression of MYC targets 1 and 2 gene class in treated vs untreated groups. AvsU, AZD5153 vs Untreated; PvsU, Pazopanib vs Untreated; CvsU, Combined vs Untreated. d Enrichment plots showing negative enrichment of hallmark MYC pathway genes in GSEA Hallmark analysis, as depicted by the profile of the running ES Score and positions of gene set members on the rank-ordered list. AvsU, AZD5153 vs Untreated; PvsU, Pazopanib vs Untreated; CvsU, Combined vs Untreated. e Representative immunoblots of PARP apoptotic marker, transcriptional regulator c-MYC, cell cycle kinase PLK1 and its downstream effectors p27 (Kip1) and p21 (Waf1/Cip1), upon treatment with AZD5153 or pazopanib, singly or in combination, for 48 h. f Real‐time polymerase chain reaction (RT‐PCR) analysis of MYC promoter regions following chromatin immunoprecipitation (ChIP) assay on LPS141 or hSC43 cells using BRD4 or rabbit IgG antibody. LPS141 cells were treated with 25 μM AZD5153 and/ or 20 μM pazopanib for 24 h while hSC43 cells were treated 80 μM AZD5153 and/ or 40 μM pazopanib for 24 h. Data presented as mean ± SD across two biological replicates. ns not significant; *, P < 0.05.