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. 2022 Jan 31;23(3):1672. doi: 10.3390/ijms23031672

Figure 5.

Figure 5

Figure 5

Effects of propolin G on GSK-3β-mediated Snail-dependent migration and invasion activities in MDA-MB-231 cells. MDA-MB-231 cells were incubated with (A) indicated concentrations of propolin G (0, 5, and 10 μM) for 24 h or (B) 10 μM of propolin G for different time intervals. The protein levels of p-Akt, Akt, p-GSK-3β, and GSK-3β were examined by Western blot. (C) The immuno-precipitation by Snail was conducted and the linked GSK-3β with Snail was evaluated by Western blot. (D) The cells were pretreated with or without MG132 (5 μM) for 1 h, and propolin G (10 μM) was added for 24 h subsequently. Snail, GAPDH, and Lamin A/C in the cell lysates of cytoplasm and nuclear fraction, respectively, were determined by Western blots. (E) MDA-MB-231 cells were stimulated with propolin G for 24 h after GSK-3β inhibitor (lithium chloride, LiCl) pretreatment for 1 h. The collected lysates were analyzed to inspect the indicated protein expressions and (F) the mRNA expression of E-cadherin. (G) The migration and (H) invasion activities of MDA-MB-231 cells were studied after lithium chloride and/or propolin G treatment as described in Materials and Methods. Data were collected from at least triplicate experiments, and the results represent the mean ± S.D. Different lowercase letters represent significant differences between the two groups when p < 0.05.