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Figure 1 | Journal of Translational Medicine

Figure 1

From: Development of a multiparameter flow cytometric assay as a potential biomarker for homologous recombination deficiency in women with high-grade serous ovarian cancer

Figure 1

Optimization of PBMC flow cytometry method. a Unstimulated PBMCs. Cells were seeded and incubated overnight without PMA/I stimulation before treatment with olaparib/carboplatin (O/C) for 48 h. bd Optimization of stimulation conditions. Condition 1: PMA/I × 4 h followed by PBS washout and subsequent O/C × 48 h (P/I × 4 h > O/C × 48 h); condition 2: PMA/I × 4 h then O/C × 24 h followed by washout and subsequent O/C × 24 h (P/I × 4 h–O/C × 24 h > O/C × 24 h); condition 3: PMA/I × 4 h followed by O/C × 48 h with no washout (P/I × 4 h–O/C × 48 h). Cells were collected after 24, 36 and 48 h of treatment before undergoing the single-stain flow cytometry protocol. b γH2AX; c MRE11; d RAD51. e, f Demonstration of optimized dual-stain. Cells were plated and treated according to condition 1 before undergoing the dual-stain process for MRE11 and γH2AX (e), or for RAD51 and γH2AX (f).

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