Figure 5. Lysosomal localization of H-ferritin in cells exposed to iron, apo-ferritin or D3T.

A) VIC was cultured in growth medium or calcification medium in the presence or absence of iron (Ammonium iron (III) citrate). Western blot analysis for H-ferritin and LAMP1 derived from the isolated lysosome. B) VIC was cultured in growth medium or calcification medium in the presence or absence of iron (Ammonium iron (III) citrate), apo-ferritin or D3T. Double immunofluorescence staining of VIC for LAMP1 and H-ferritin are shown. Images were obtained employing immunofluorescence-confocal microscope and STED nanoscopy. Representative staining is shown from at least three independent experiments. Results were analyzed by One Way ANOVA, Bonferroni’s Multiple Comparison Test and are shown as mean values ± SEM of at least three independent experiments. ***P < 0.0001.