Fig. 8: FIP1 overexpression leads to global activation of CPA activity. | Nature Communications

Fig. 8: FIP1 overexpression leads to global activation of CPA activity.

From: Elevated pre-mRNA 3′ end processing activity in cancer cells renders vulnerability to inhibition of cleavage and polyadenylation

Fig. 8: FIP1 overexpression leads to global activation of CPA activity.The alternative text for this image may have been generated using AI.

a Schematic of a lentiviral vector containing an inducible FIP1 (top) and Western blot (bottom) showing induction of FIP1 in HEK293T cells by 2 μg/mL Dox for one or two days. b Scatter plot comparing 3′UTR isoform abundance difference between HEK293T cells with FIP1 induction (Dox+, 24 h) and those without (Dox−). Each dot represents a gene with two selected 3′UTR isoforms (as shown in Fig. 5a). c As in b, except that IPA isoforms are analyzed. See Fig. 5f for IPA and TPA isoform definitions. d Schematic of reporter assay to examine APA regulation by FIP1 overexpression (OE) using pTRE-FIP1-IRES-BFP. e Bar graph indicating the level of pPAS usage in HEK293T cells with the blue fluorescent signal tracking FIP1 expression level. Cells showing no, low, or high blue signals were analyzed for pPAS usage, as indicated by log2(R/G). Error bars are standard deviation based on three biological replicates. P values (t-test) for significance of difference between comparing groups are indicated. f Inhibition of pPAS usage by JTE-607 in cells with different levels of FIP1 OE. Linear regression formula and R2 values are shown for the cell groups with no or high BFP signals. Error bars are standard deviation of three biological replicates. P value (t-test) for significance of difference in Δlog2(R/G)(10 μM JTE-607 vs. DMSO) between cell groups are indicated. g Schematic of the cell competition assay, in which HepG2 or HeLa cells with inducible FIP1 OE (named HepG2/iFIP1 and HeLa/iFIP1, respectively) are subject to JTE-607 treatment, followed by flow cytometry analysis to examine green or blue fluorescent signals of surviving cells. h Histograms showing distribution of fluorescence signals of HepG2 cells after DMSO (top) or JTE-607 (bottom) treatment. i Bar graph showing ratio of number of cells with low (bottom 1/3) to high fluorescent signals (top 1/3) in HepG2 cells treated with 1 μM JTE-607. Error bars are standard deviation of three biological replicates. P (t-test) for significance of difference is indicated. j As in i, except that HeLa/iFIP1 cell data are shown with 50 μM JTE-607. Source data are provided as a Source Data file.

Back to article page