Fig. 1.

Alkyl (lipid peroxyl) radical generation in cultured podocytes. (A) Conditionally immortalized mouse podocytes were cultured as described in Methods. Representative pictures show podocytes maintained at 33 °C with IFNγ present and for differentiation cells were switched to non-permissive conditions (37 °C, no IFNγ). Podocytes were differentiated for at least 11 days or until increased synaptopodin levels were evident as verified by Western blot analysis. (B) The free radical donor AAPH was used to generate alkyl radicals and mimic lipid peroxidation. Equations show the temperature sensitive decomposition of AAPH, the resultant radical formation in the cell membrane and the method for calculating free radical production for the incubation periods.