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. 2020 Apr 27;11(6):1594–1602. doi: 10.1111/1759-7714.13435

Figure 3.

Figure 3

Effects of PDGF‐BB treatment on the combination of Cx43, AKAP95, and cyclin E1/E2. (a) Starvation treatment with serum‐ and ‐antibiotic‐free medium for 24 hours was employed when the cell density reached 50%–60%. The cells were then treated with a medium containing series concentration gradient of PDGF‐BB of 5, 10, 20, and 40 ng/mL for 24 hours. The control group cells were cultured under normal conditions synchronously. The total protein was extracted to detect the expression of AKAP95, Cx43, cyclin E1, and cyclin E2 by western blot analysis. (b) Starvation treatment with serum‐ and ‐antibiotic‐free medium for 24 hours was employed when the cell density reached 50%–60%. The cells were then treated with a medium containing 15 ng/mL PDGF‐BB for 24 hours. Total protein of 500 μg extracted from the experimental and control groups was taken. The operation and process to detect Cx43, AKAP95, cyclin E1 and cyclin E2 were similar to that in Fig 2b. The ratio of PDGF‐BB treatment group to control group was taken as a statistical variable for comparison. *P < 0.05; **P < 0.01. (c) Starvation treatment with serum‐ and antibiotic‐free medium for 24 hours was employed when the cell density reached 50%–60%. The cells were then treated with a medium containing 15 ng/mL PDGF‐BB for 24 hours. Total protein of 500 μg extracted from the experimental and control groups was taken to detect the expression of kinases with antibodies ERK1/2, ERK5, PKA, PKB and PKC using western blot analysis. *P < 0.05; **P < 0.01.