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. 2014 Sep 22;3(9):e119. doi: 10.1038/oncsis.2014.33

Figure 4.

Figure 4

The MAPK cascade mediates PMA-induced MMP-9 proteolytic activity in cells deficient for FLNB. (a) Representative immunoblots detecting protein levels of FLNB, RAS-GRF, phosphorylated PKC-α/β, phosphorylated PKC-δ/θ, total PKC-δ, phosphorylated ERK1/2, total ERK1/2 and FLNA in serum-starved Flnb+/+ and Flnb−/− MEFs after induction with or without PMA for 15 min. Actin served as an internal loading control. (b) Representative immunoblot demonstrating the inhibition of p-ERK1/2 expression in Flnb−/− cells induced with PMA and pretreated with PD 098059, a MAPK inhibitor. *P<0.05 versus non-pretreated MEF with PD 098059. (c) Gelatin zymography detecting proteolytic activity of MMP-9 in cell culture medium of Flnb−/− MEFs pretreated with PD 098059, and then cultured in serum-starved conditions with PMA. (d) Immunoblots for pro-MMP-9, MMP-9, p-ERK1/2 and total ERK1/2 proteins extracted from Flnb−/−H-RAS+ and Flnb+/+H-RAS+ tumors. Actin served as an internal loading control. (e) Densitometric reading of MMP-9 and p-ERK1/2 proteins are given as fold differences. Data of triplicate or quadruplicate experiments are expressed as mean±s.d. *P<0.05; ***P<0.001 versus non-PD 098059-treated MEFs or Flnb+/+H-RAS+ tumors.