Fig. 3: Profiling of BE(2)N cells during BrdU-induced switch from ADRN to MES cell types.

a Morphology of BE(2)N at different time points following BrdU treatment. (b) TRAP analysis of telomerase activity in BE(2)N at different time points during BrdU-induced phenotypic switch. The assays were performed using serial dilutions of the extracts (equivalent to 500, 50 and 5 ng of proteins), and the relative activities (mean ± S.D., n = 2 or 3 experimental replicates) are plotted at the bottom. Pair-wise P-values were determined using student’s t-tests (two-tailed). c Western analysis of telomere and lineage-related proteins in BE(2)N during BrdU treatment. d Analysis of telomere length distributions in BE(2)N at different time points during BrdU-induced phenotypic switch. e The number of upregulated and downregulated genes in BrdU-treated BE(2)N (by >2-fold in comparison to the DMSO-treated control cells) were determined from RNA-seq analysis and plotted. f The expression patterns of the ~1100 genes that show the greatest changes in mRNA levels (>3 fold) in BrdU-treated BE(2)N were displayed using Heatmap. g The list of genes that were downregulated in BrdU-treated BE(2)N (by >2-fold) was compared to the ADRN signature list¹⁰. In parallel, the list of genes that were upregulated in BrdU-treated BE(2)N (by >2-fold) was compared to the MES signature. h The ~2000 genes upregulated in BrdU-treated BE(2)N (by >2-fold) were subjected to pathway analysis by the Enrichr program. The top ten GO terms identified by the KEGG pathway database and the Hallmark gene sets in the Molecular Signature Database were ranked by adjusted P value and plotted.