Figure 2.

TET-regulated expression of EGFP-HA-ferritin in C6 rat glioma cells augments iron uptake. (a) C6-TET-EGFP-HA-ferritin cells (clone 2) were grown in culture plates in the presence or absence of TET (48 hours; 1 µg/ml). Confluent cell layers were examined for EGFP fluorescence (upper panel) and in combination with bright field (lower panel). Scalebar = 50 µm. (b) Time-dependent expression of EGFP and HA-ferritin in C6 cells as a response to TET switch (1 µg/ml) was determined by Western blot analysis. β-Actin was used to normalize protein amounts. (c) Prussian Blue analysis of dose-dependent TET-induced (48 hours) iron uptake into the cells. Staining intensity was quantified by NIH image. (d) Quantification of intracellular iron content by ICP-AES (1 µg/ml; 48 hours). Close and open bars represent clones 1 and 2, respectively.