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. 2023 Feb 3;14:970457. doi: 10.3389/fphar.2023.970457

FIGURE 1.

FIGURE 1

Influence of palbociclib and fulvestrant on T cell proliferation. (A) eFluor™ 670-stained T cells were stimulated by anti-CD3/CD28 beads and cultured in the presence or absence of palbociclib (daily addition), fulvestrant, or their combination at indicated concentrations for 24, 48, 72, 96, 120, 144, and 168 h. Cells were harvested and dilution of eFluor670™ dye was determined by flow cytometry. The results are depicted as the means ± SEM of four donors. (*p ≤ 0.05 compared to control sample “stim + DMSO”). (B,C) eFluor™ 670-stained T cells were stimulated by anti-CD3/CD28 beads and cultured in the presence or absence of palbociclib (single administration), fulvestrant, or their combination for (B) 48 h and (C) 120 h. Cells were harvested and dilution of proliferation dye eFluor670™ was determined by flow cytometry. The results are depicted as the means ± SEM of three donors. (*p ≤ 0.05 compared to control sample “stim”). (D) DNA replication in proliferating cells was analyzed by EdU flow cytometry assay. Therefore, T cells were stimulated by anti-CD3/CD28 beads and cultured in the presence of palbociclib (daily addition) at indicated concentrations for 96 h. 18 h prior to cell harvesting, cells were incubated with 10 µM EdU. Cells were harvested and percentage of EdU-positive cells was determined by flow cytometry. (*p ≤ 0.05 compared to control sample “stim + DMSO”).