Reduced PIKfyve lipid products in Vps34 KO podocytes. Vps34fl/fl podocytes, seeded on 35 mm dishes, were treated with Ad-Cre or Ad in complete culture media, as described under “Materials and Methods”. On day 4 post-treatment media was replaced with inositol/glucose-free media for 24 h, followed by myo-[2-3H]inositol labeling for 26 h. Lipids were extracted, deacylated and fractionated by HPLC. Shown is quantitation from three independent experiments, calculated as a percentage of the total PI radioactivity and presented as mean ± SE. The total PI radioactivity was obtained by summing the counts within the elution times corresponding to the indicated [3H]GroPIns peaks. *, PtdIns(3,5)P2, PtdIns5P and PtdIns3P in Vps34 KO vs. those in Vps34fl/fl podocytes, P<0.05.