Characterization of YOD1-C160S transgenic mice. (A) YOD1-C160S mice were fed with normal or doxycycline (2 mg/mL) supplemented water. Lysates of indicated organs from euthanized mice were resolved by electrophoresis and immunoblotted using an anti-HA antibody. The top panel shows the kinetic of YOD1-C160S expression in mice on doxycycline for 2, 5, or 8 days. The bottom panel shows expression of YOD1-C160S in different organs as indicated at 8 days of doxycycline supplementation. (B) HA immunoblots show expression of YOD1-C160S (top panel) and total YOD1 (middle panel, arrows: YOD1-C160S, wedge: endogenous YOD1, asterisk: background band) in BMDCs on day 7. Anti-PDI blot served as loading control. (C) YOD1-C160S mice were given doxycycline-water for 7 days and splenocytes were isolated thereafter. Intracellular staining in surface stained CD11c⁺ and CD11b⁺ cells was performed using Alexa Fluor 488–conjugated anti-HA antibody. (D) Intracellular staining for YOD1-C160S expression in control and doxycycline supplemented BMDCs using anti-HA Alexa Fluor 488 conjugated antibody is shown. (E) Mice were given doxycycline water for ∼ 20 days and cellular distribution in the lymphoid organs of control and doxycycline supplemented mice is shown by representative FACS plots.