TIMP3-induced apoptosis is influenced by serum factors.

<p><b>A:</b> Influence of TIMP3 on activation of cRaf, ERK1/2, RSK1 and Akt was determined in Cal78 cells and transduced Cal78 cells with LacZ or TIMP3 by spot array measurements. <b>B:</b> Phosphorylation of cRaf, ERK1/2, RSK1 and Akt was confirmed by western blotting. <b>C:</b> Influence of serum withdrawal on apoptosis rates after 24 hours. Apoptosis in lentiviral transduced cells was measured relative to Cal78 cells cultured with 10% FCS. Values less than p<0.05 (*or °) were considered statistically significant. <b>D:</b> Influence of specific growth factors on apoptosis rates under serum-free conditions. Prior to the assessment of apoptosis, Cal78 cells and transduced Cal78 cells with LacZ or TIMP3 were stimulated with 100 ng/ml EGF, TGF-ß or FGF-2 and cultured for 24 hours. Apoptosis was defined by caspase 3 and 7 activities. *Indicates statistical significance (p<0.05). <b>E:</b> Influence of serum withdrawal on autophagocytosis in Cal78 cells after 24 hours. Autophagocytosis in TIMP3 overexpressing CAL78 cells was determined relative to Cal78 cells transduced with a control construct (LacZ) cultured with 10% FCS, 1% FCS or ITS by Western blot analysis of the typical marker proteins LC3 and Beclin.