Fig. 1: Biochemical and cellular activity of AB25583.
From: Structural basis for a Polθ helicase small-molecule inhibitor revealed by cryo-EM
a Structure of AB25583. b Schematic of Polθ-hel ATPase activity assay. c Scatter plot showing AB25583 inhibition of the indicated SF2 DNA helicases. Data represent the mean of two technical replicates. IC50 of AB25583 against Polθ-hel = 6 nM. d Scatter plot showing AB25583 inhibition of Polθ-hel ATPase activity in the presence of the indicated ATP concentrations. Data represent the mean of two technical replicates. e, f, l, o Statistical significance was measured from a two-sample t-test and P values are indicated. e, f Scatter plot showing % colony survival in the presence of the indicated concentrations of AB25583. Data represent the mean of three biological replicates. n = 3, ±s.e.m; P = 0.002507 for 0.5 μM and P = 0.000044 for 1 μM in HCT pair; P = 0.00278 for 0.5 μM and P = 0.000245 for 1 μM in DLD1 pair. g Structure of AB25595. h Scatter plot showing AB25595 inhibition of Polq-hel. Data represent the mean of three biological replicates. n = 3, ±s.d. i–l Scatter plot showing % colony survival of the indicated cell lines in the presence of the indicated concentrations of AB25595 (i) or AB25583 (j–l). i, k, l Data represent the mean of three biological replicates. n = 3, ±s.e.m; P = 0.00185 for 10 uM for Polq −/− vs sgGFP. j Data represent the mean of two biological replicates. n = 2, ±s.e.m. m Bar plot showing % nuclei with >5 gH2AX foci following treatment with the indicated concentrations of AB25583 (right). Data represent the mean of three biological replicates. n = 3, ±s.e.m. Representative images of gH2AX immunofluorescence following DMSO and AB25583 treatment Magnification 40x; Scale bar, 10 um (left). P = 0.00303 for 5 μM and P < 0.00001 for 10 μM in BRCA2-KO. n Bar plot showing % nuclei with >5 RAD51 foci following DMSO and AB25583 treatment. Data represent the mean of three biological replicates. n = 3, ±s.d. P = 0.019585 for IR treated, DMSO vs AB25583. o Bar plot showing quantitation of MMEJ in U2OS cells indicated by % of GFP/dsRED following DMSO and 20 μM AB25583 treatment. Data represent the mean of three biological replicates. n = 3, ±s.d. P < 0.0001 for DMSO vs 20 μM AB25583. Source data are provided as a Source data file.
