Fig. 2

CDK9i transiently suppresses expression of oncoproteins. A OCI-LY3 and VAL cells treated with 30 nM AZD4573 or vehicle control for 3 h and subjected to proteomic analysis. Heatmap of all detected proteins is shown. Data are represented as z scores calculated from normalized protein abundance. B Volcano plot of all differentially expressed proteins common to both cell lines (|FC| ≥|1.5| treatment versus control; p ≤ 0.05). The identities of select proteins are shown. C Cell lines were treated with AZD4573 (30 nM) for 0, 3 and 8 h prior to harvest. After 8 h exposure, the compound was either washed out (w) or not (c = continuous exposure) and harvested after 24 h. Whole cell lysates were subjected to immunoblotting. D Top enriched and depleted pathways from IPA analysis of proteomics data from AZD4573-treated VAL and OCI-LY3 cells. Data is presented as a dot plot ranked by pathway Z-score, with size representing the number of genes and color indicating the -Log10 of the significance. E VAL (blue) and OCI-LY3 (red) cell were treated with AZD4573 for 0, 3 and 8 h prior to harvest. After 8 h exposure, the compound was washed out and cells were harvested after 24 h. mRNA expression of select genes was quantified by RT-PCR. Data is shown as mRNA fold change in cells treated with AZD4573 versus time-matched cells treated with DMSO. Bars represent mean ± SEM of three independent experiments. Note that plots for the gene PIM3 were separated from the other genes due to the higher y-axis scale. *p < 0.05 and **p < 0.01, AZD4573 vs. time-matched DMSO control