Rpgrip1l Knockout Mouse Model and Cell Experiments

The Rpgrip1l mouse model was described previously [2 (link),4 (link),5 (link),25 (link)]. HaCaT cells were transfected with RPGRIP1L siRNAs (HSC.RNAI.N015272.12.1 and 2, Integrated DNA Technologies, Coralville, IA). Non-targeting (Negative Control) siRNA (NC-1, Integrated DNA Technologies) was used as control siRNA. Twenty-four hours after transfection, cells were switched to high calcium (1.5 mM CaCl₂) for designated durations. EGTA was used at 2 mM. Dynasore (Sigma-Aldrich, Saint Louis, MO) and sucrose were used at 50 μM and 400 mM, respectively. PV IgG were purified in Dr. Payne’s laboratory and used at 400 μg/ml. Normal human IgG (Sigma-Aldrich, Saint Louis, MO) was used as control IgG. Method details are provided in the Supplementary Materials and Methods (S1 Text) online.