Lentiviral infection and production

All shRNA plasmids were purchased from OriGene, Rockville, MD, already cloned into the pGFP-C-shLenti backbone: RapGEF2 shRNA plasmids (TL316875), MAGI2 shRNA plasmids (TL311600), and scrambled shRNA control (TR30021). Hairpin sequences specific for human RapGEF2 are as follows: CAATGTCAGTGAGGCGA GAACTCTGTGCT (#1) and TGGTCAGTCTCAAGATGACAGCA TAGTAG (#2). Hairpin sequences specific for human MAGI2 are as follows: GAACCTGAGCCATACAGAAGTAGTGGATA (#1) and TCTACTTCATTGACCATAACACAAAGACA (#2). All lentiviral preparation and infections were performed as previously described.²⁹ Infections were done at the permissive temperature in conditionally immortalized human podocytes, and then stable cell lines were established by selection with puromycin at a concentration of 1 to 2 ug/ml. Knockdown and control transduced cell lines were grown at 37 °C for at least 1 week prior to use in experiments.