Fig. 4

The mRNA expression of DR4, DR5, p53, and TRAIL genes in these cells over time, along with the assessment of the apoptotic proteins involved in PANC-1 cells following a 72-hour treatment with ONC201, TBP-134, and TBP-135. The normalised expression of (A) DR4, (B) DR5, (C) p53 and (D) TRAIL genes are shown in column charts. (E) The proteome profile array blots of PANC-1 after treatment with 0.5 µM ONC201, TBP-135 and TBP-134 for 72 h. The dots for each protein are duplicates. The green squares indicate the pro-apoptotic proteins, while the red squares show the anti-apoptotic proteins quantified on the bar graphs. The relative (F) pro- and (G) anti-apoptotic protein levels are shown in bar charts. (H) Images of PANC-1 cells, including merged brightfield and fluorescence (FITC) channels as well as fluorescence (FITC) channel images, were captured using the Celldiscoverer 7 after 72 h of treatment with ONC201 and TBP-135. The green dots indicate the caspase 3/7 active cells. The abbreviation B + F merged refers to the merged image of the Brightfield and Fluorescence (FITC) channels, while Fluor. channel represents the Fluorescence (FITC) channel alone and M stands for Medium control. (I) The number of Caspase-3/7 active cells shown in column charts after 72 h of treatment with ONC201 and TBP-135. The red lines indicate the threshold of the changes in expression. The data were normalised to the DMSO control. The data are presented as mean values ± standard deviation (SD) (n = 3 – gene expression, n = 2 – proteom profile, n = 1 – caspase 3/7 activity). Significance levels are shown as x - p < 0.05; y - p < 0.01; z - p < 0.001 using one-way ANOVA test followed by Fisher’s LSD post hoc test.