Fig. 3.

Patient fibroblasts show reduced proliferation and impaired S-phase entry. A Representative images of skin-derived primary fibroblasts from a healthy control and the patient, 1 day and 7 days after seeding. Images were acquired with an EVOS7000 microscope, original magnification × 4. B Fibroblast proliferation in healthy controls and patient. Cell counts were performed with ImageJ. Data are representative of 2 independent experiments. C Representative FACS plots and histograms of cell-cycle analysis of healthy control and patient primary fibroblasts upon synchronization at G1 and release at 0 and 24 h. Data are representative of 3 independent experiments. D Percentage of cells in G1, S, and G2 phase in healthy controls compared to the patient, upon synchronization at G1 and release at 0 and 24 h. E Representative FACS plots from S phase entry and progression analysis of healthy control and patient primary fibroblasts upon synchronization at G1 and release. Nocodazole was used in the release medium to inhibit mitosis and block cells in G2 phase. Data are representative of 3 independent experiments. F Percentage of cells in G1, S, and G2 phase in healthy controls and the patient, upon synchronization at G1 and release for 0, 24, 48 h. Results represent means ± SEM