Figure 2.

Effect of abemaciclib on mesothelioma 3D cultures. (A) DPM cell lines (NCI-H28, NCI-H2452, MSTO-211H, and NCI-H2052) are represented as 2D monolayer (left) and 3D single spheroid. Images were acquired 4 days after seeding. (B) DPM single spheroids were treated with abemaciclib on day 0. Cells were treated with abemaciclib (5 µM) or DMSO, as a control, starting on day 0. Images were acquired on days 0, 1, 2, 3, and 4. Abemaciclib at low concentration (5 µM) significantly inhibited the ability of DPM cells to generate spheroids. All experiments were performed at least three times. (C) DPM 2-day-old spheroids were treated with abemaciclib. Cells were seeded at a density of 0.5 × 10⁴ cells/well in U-bottom ULA 96-well plates and after 2 days were treated with abemaciclib (10 µM) or DMSO as control. Images were acquired on days 0, 1, 2 (day of treatment), 3, and 4. Experiments were performed at least three times. (D) Cells were seeded at a density of 30 × 10⁴ cells/well in ULA 6-well plates and after 2 days treated for 72 h with abemaciclib or DMSO as control. Spheroids were disaggregated in single cells and seeded again. Images were acquired on days 0, 1, and 2 (day of treatment); day 5 for first-generation spheroids; and days 1, 2, and 3 for second-generation spheroids. All experiments were performed at least three times. DPM, diffuse pleural mesothelioma; DMSO, dimethyl sulfoxide.