Figure 4.

Transfection with FH- or FLsiRNA altered the levels of ferritin chains as determined by immunolocalization. The cells were transfected with control nontargeting siRNA (A, D, G, J), FHsiRNA (B, E, H, K, M–O), or FLsiRNA (C, F, I, L) and immunolabeled after 4 days for ferritin chains using (A, B, C) H- or (D–F, J–M) L-specific antisera followed by Alexa Fluor-488-conjugated secondary antibodies (green fluorescence) and DAPI staining (blue nuclei). Negative antibody controls were incubated with normal rabbit serum (Control serum) in place of ferritin-specific antisera (G, H, I). Higher magnifications of representative cells for each siRNA treatment and labeled for L-ferritin are shown (J–L). (M) A cell transfected with FHsiRNA for 4 days and labeled for 1 hour with red fluorescent dye before fixation (N). The overlay (O) shows colocalization (yellow) of ferritin within lysosomes (arrows).