Ab2302

Rat lung Paraffin sections (5 μm thick) were obtained following in vivo experiments as previously described [19 (link)]. Sections were incubated with peroxidase blocking solution (Dako, Cambridge) and then with primary antibodies for rabbit anti-active caspase-3 (1:50 Abcam ab2302), NF-κB p65 (1:200 Cell Signaling C22B4), rabbit anti-phospho-IKKα/β (1:40 Cell Signaling 2697), P-Stat3 (1:50 Cell Signalling 9145); Stat3 (1:400 Cell Signalling 9149) or SMA (1:400 Dako M0851). Sections were then incubated with polyclonal goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody (Dako) followed by incubation with diaminobenzidine (DAB) and peroxide buffer (Sigma) to produce a brown stain. Slides were counterstained with hematoxylin or eosin to provide nuclear and morphological detail. Non- specific rabbit IgG (Sigma-Aldrich) at the same concentration as those used above was used as a control.

The prostatic cancer cell line LNCaP was obtained from ATCC (American Type culture collection, USA) and maintained in our laboratory for the study. Cells were cultured in RPMI 1640 (Gibco™, 32404-014), supplemented with 10% fetal bovine serum (Gibco™, 10500-064) with or without 4mM L-glutamine (Gibco™, 25030-081), 100 U/ml of penicillin and 100ug/ml streptomycin (15140122, Life Technologies) at 37°C, and 5% CO2. Anti-PDH antibody (cell signaling, C54G1), anti-Pyruvate Dehydrogenase E1-alpha subunit antibody (ab110334), monoclonal anti-GAPDH antibody (sigma, AB2302), anti-GLUT1 antibody (cell signaling, D3J3A), Anti-LDHA antibody (abcam, ab47010), anti-HK2 (abcam, ab104836), anti-GLS1 (sigma, WH0002744M1-100UG), anti-GLUD1 (Novus Biologicals, NBP1-54961), anti-MPC1 (Novus Biologicals, NBP1-91706) and anti-MPC2(abcam, ab111380) were applied in the current study. BPTES (SML0601), EGCG (03970590-10MG) and dimethyl-αKG (34963-1) were obtained from Sigma-Aldrich (Sigma-Aldrich Norway AS Oslo, Norway). Annexin-V was purchased from Life technologies (Life technologies, A13199).