Recombinant γ interferon

Pkd1-depleted mouse kidney tubule cell lines (Pkd1^+/- and Pkd1^-/- cells) (a kind gift from Dr. Changlin Mei of Changzheng Hospital, Second Military Medical University, Shanghai, China) were cultured in Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (DMEM/F12) (Gibco) supplemented with 2% fetal bovine serum (Gibco), Insulin-Transferrin-Selenium (41400045, Gibco), triiodothyronine (2 x 10^-9 M, T5516, Sigma) and recombinant γ-interferon (10 units/ml, Sigma) at 33°C. For cilia formation, cells were transferred to 37°C and grown in γ-interferon-free medium for 7 days.

Mouse inner medullary collecting duct (mIMCD3) cells were obtained from ATCC and grown in DMEM low glucose medium supplemented with 10% FBS and maintained in 5% CO₂ atmosphere at 37 °C. Pkd2^−/− cells were obtained from Steve Somlo and grown in DMEM:F-12 medium (catalogue # 10565-018,Gibco) supplemented with 2% foetal bovine serum, insulin (8.3 × 10⁻⁷ M), prostaglandin E1 (7.1 × 10⁻⁸ M), selenium (6.8 × 10⁻⁹ M), transferrin (6.2 × 10⁻⁸ M), triiodothyronine (2 × 10⁻⁹ m), dexamethasone (5.09 × 10⁻⁸ m) and recombinant γ-interferon (10 units/ ml⁻¹, Sigma) at 33 °C50 (link) Human primary ADPKD cells were grown in DMEM:F-12 medium (catalogue # 10565-018,Gibco) supplemented with 5% FBS, 5 μg ml⁻¹ insulin, 5 μg ml⁻¹ transferrin and 5 ng ml⁻¹ sodium selenite (ITS)(catalogue # 17-838Z, Lonza)50 (link).