Extended Data Fig. 8: Validation of the sensitivity of PPP2R1A-mutant tumours to immunotherapies in pre-clinical models.
From: PPP2R1A mutations portend improved survival after cancer immunotherapy
a. Reduced PPP2R1A expression by gene-specific shRNAs. Three independent pairs of PPP2R1A-specific shRNAs (sh#1-3) were used to generate a set of SKOV3 cell lines with PPP2R1A-knockdown (KD). SKOV3 transduced with the non-targetable shRNA (shNC) was used as the negative control. Two independent experiments were performed, with the representative results shown. For gel source data, see Supplementary Fig. 1. b-c. Generation of SKOV3 cell lines with stable overexpression of PPP2R1A WT and P179R mutant. SKOV3 transduced with lentiviral vectors encoding PPP2R1A WT and P179R mutant were selected by sorting GFP positive cells. SKOV3 transduced with empty vector was used as the negative control. d. Flow sorting of parental and CRISPR-edited HEC50B and OVCAR429 cell lines. Enriched CD19+ cancer cells were subsequently used for cytotoxicity assays. e. Validation of PPP2R1A mutational status in PDX by Sanger-sequencing. f-g. Humanized-BLT mice bearing PPP2R1A-WT endometrial cancer PDXs were randomized into the indicated treatment groups. Images of tumours from the indicated groups are shown at the end of treatment (f). Tumour weights were measured as surrogate for tumour burden (g). Error bars represent mean ± s.e.m. For panel g, P values were calculated using two-sided Student’s t-test. h. Validation of Ppp2r1a mutation status in mouse OCCC cell lines applied in syngeneic models. i. Mouse body weights measured during treatment in the Arid1a−/− Pik3caH1047R and Ppp2r1aR183Q Arid1a−/− Pik3caH1047R syngeneic mouse OCCC models.
