Metformin and IR reduce microvessels and enhance apoptosis markers in A549 LC tumours. (A) Effects on apoptosis markers. Lysates from control, MET-, IR- and MET+IR-treated tumours were analysed with immunoblotting using anti-CD31, -Puma, -Bax and -actin antibodies. Representative immunoblots are shown. (B) Representative images of anti-CD31 IHC analysis of A549 control (CON), MET-, IR- and MET+IR-treated tumours. (C) Densitometric analysis of immunoblotting results from lysates of tumours in all experimental groups (n=6 in each group). Data are mean±s.e. of normalised densitometry values (*, P⩽0.05; **, P⩽0.001 statistically significant difference between treatment groups and control for each marker). (D) Effects MET and IR on expression of CC3. Representative images of IHC analysis of control (CON), MET-, IR- and MET+IR-treated tumours with an anti-CC3 antibody.