Extended Data Fig. 4: Validation of Genes that Modulate Response to Cyclin A/B RxL Inhibitors or CDK2 Inhibitors.
From: Targeting G1–S-checkpoint-compromised cancers with cyclin A/B RxL inhibitors
a-b, Dose response assays of NCI-H1048 cells infected with two independent non-targeting sgRNAs (sgCtrl) or two independent sgRNAs against LIN54 (a) or ZWINT (b), and treated for 6 days with increasing doses of the cyclin A/B RxL inhibitor (CIRc-004). c-f, Dose response assays of NCI-H446 cells infected with sgCtrl or sgRNAs against C) CCNB1 (c), CDK2 (d), KNTC1 (e), MAD1L1 (f), treated for 6 days with increasing doses of CIRc-004. g-h, Dose response assays of NCI-H1048 cells infected with two non-targeting sgRNAs (sgCtrl) or two independent sgRNAs against CCNB1 (g) or CDK2 (h), and treated for 6 days with increasing doses of the Cdk2 inhibitor (PF-07104091). i) Representative flow cytometric analysis of phospho-histone H3 in NCI-H1048 cells infected with the indicated sgRNAs and treated with CIRc-004 at 20 nM or DMSO for 24 h. j-k, Quantitation of cells in mitosis by flow cytometry analysis using phospho-histone H3 from i (j) or PI (k) of NCI-H1048 cells transduced with the indicated sgRNAs and then treated with CIRc-004 at 20 nM or DMSO for 24 h. l) Representative flow cytometric analysis from Fig. 2h of cleaved PARP in NCI-H1048 cells infected with the indicated sgRNAs and treated with CIRc-004 at 20 nM or DMSO for 3 days. For i, data are shown as fold change relative to vehicle. For a-h, j, k, n = 3 biological replicates and data are mean +/– SD. Arrows in a-h indicates DMSO-treated sample used for normalization. Statistical significance in j, k was calculated using unpaired, two-tailed students t-test. Where indicated, *=p < 0,05, **=p < 0.01.
