HK2 cells were obtained from ATCC (Manassas, VA) and were cultured with Keratinocyte Serum Free Medium (K-SFM) supplemented with 0.05mg/ml BPE and 5ng/ml EGF (Invitrogen), without added glucose. Cells were seeded at a density of × 105 per well and treated for 72 hrs with glucose (30mM), IL-6 (1nM), leptin (5nM), FGF21 (10nM), IL-1β (1nM) or TGF1β (400pM) (Abcam, USA). A total of 4–10 different replicates were used for each condition.
NCI-H295R cells were obtained from ATCC (Manassas, VA) and cultured in DMEM:F12 medium, supplemented with 0.00625mg/ml insulin, 0.00625mg/ml transferrin, 6.25ng/ml selenium, 1.25mg/ml bovine serum albumin, 0.00535mg/ml linoleic acid and 2.5% Nu-Serum (Corning, USA). Cells were seeded at a density of × 105 per well and treated for 24 hrs with 100ng/ml IL-6 (1nM), leptin (5nM), FGF21 (10nM), IL-1β (1nM), TGF1β (400pM) or IL-10 (1nM) (Abcam, USA). A total of 4–10 different replicates were used for each condition.
NCI-H295R cells were obtained from ATCC (Manassas, VA) and cultured in DMEM:F12 medium, supplemented with 0.00625mg/ml insulin, 0.00625mg/ml transferrin, 6.25ng/ml selenium, 1.25mg/ml bovine serum albumin, 0.00535mg/ml linoleic acid and 2.5% Nu-Serum (Corning, USA). Cells were seeded at a density of × 105 per well and treated for 24 hrs with 100ng/ml IL-6 (1nM), leptin (5nM), FGF21 (10nM), IL-1β (1nM), TGF1β (400pM) or IL-10 (1nM) (Abcam, USA). A total of 4–10 different replicates were used for each condition.