Anti dgkα

Anti‐DGKα (Cat# H00001606‐B01P), anti‐CD204 (Cat# PAB30252), and anti‐Gli1 (Cat# MAB16740) antibodies were provided by Abnova. Anti‐FAK (Cat# 39−6500) antibody was obtained from Invitrogen. Antibodies against CCL22 (Cat# ab217351) and CCR4 (Cat# ab83250) were obtained from Abcam. Antibodies against AKT (Cat# 9272) and SuFu (Cat# 2522) were obtained from CST. Antibody against Lamin A/C (Cat# A17319) was obtained from ABclonal. pGli1 Ser¹¹²/Thr¹¹⁵/Ser¹¹⁶ antibody was synthesized from ABclonal.

Antibodies to p-Src Y416, Src (clones 36D10 and L4A1), p-rpS6 S240/4, cyclin D3, cleaved-caspase 3 and myc-tag (clone 9B11) were purchased from Cell Signaling. Anti-DGKα was from Abnova, monoclonal anti-β-actin and -α-tubulin antibody from Sigma-Aldrich, and anti-GAPDH from Santa Cruz Biotechnology. Anti-phosphotyrosine (4G10) was from Upstate.
Leupeptin and aprotinin were from Roche, growth factor reduced-matrigel from BD Biosciences. R59949 (DGK inhibitor II; 3-{2-(4-[bis-(4-fluorophenyl) methylene]-1-piperidinyl)ethyl}-2,3-dihydro-2-thioxo-4(1H)quinazolinone), PP2, LY294002 and Akt Inhibitor VIII (Akti) were from Calbiochem. All other reagents were from Sigma-Aldrich.

Antibodies to p-S6K Thr389 (9205), S6K (9202), p-AKT Ser473 (9271), AKT (2920), p-S6 Ser240/4 (2215), S6 (2317), p-PKD Ser744/8 (2054) and p-PKC substrate (2261) were from Cell Signaling (Danvers, MA, USA), anti-PKCα Ser657 from US Biologicals (Swappscott, MA, USA) (P9103-16A), anti-PKCα and PKD from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (sc-208, -935, -935), anti-SREBP-1 from Millipore (Temecula, CA, USA) (04-469), anti-DGKζ from Abcam (Cambridge, UK) (ab105195), anti-DGKα from Abnova (Tapei, Taiwan) (BO1P), anti-tubulin DM1A (T9026) from Sigma-Aldrich (Saint Louis, MO, USA) and anti-phosphotyrosine (4G10) from Upstate (Lake Placid, NY, USA). Horseradish peroxidase-coupled polyclonal goat anti-mouse and -rabbit immunoglobulins were from DakoCytomation (Glostrup, Denmark) (P0447, P0448). DGK inhibitor II (R59949; 266788), bisindolylmaleimide and Gö6976 were from Calbiochem (Darmstadt, Germany), (203291, 36520). Rapamycin was from LC Laboratory (Woburn, MA, USA) (R-5000). Torin was from Tocris (Bristol, UK) (4247). Recombinant DGK from Escherichia coli (D3065), PMSF, leupeptin and aprotinin were from Sigma-Aldrich. Adenosine 5′triphosphate [γ³²P] was from Hartman Analytic (Braunschweig, Germany). All lipids were purchased from Avanti Polar Lipids (Alavaster, AL, USA). Organic reagents were from Merck (Darmstadt, Germany).