Figure 9.

BSA decreases intracellular porphyrin accumulation after ALA + DFO treatment of HuH-7 cells. HuH-7 cells were treated with ALA + DFO (A/D) for 16 h in serum-free media (supplemented with 5 mg/ml BSA where indicated). Porphyrins were analyzed in the cells and the media by UPLC. A and B, UPLC chromatograms show the amounts of fluorescent porphyrins inside the cells (A) and in the media (B). Copro and PP-IX peaks are labelled, and arrowheads indicate unidentified fluorescent products that are formed after ALA + DFO treatment. C, quantification of Copro and PP-IX levels calculated from the area under the curve of the fluorescent peaks shown in panels A and B. The data are average of three independent experiments with error bars representing the standard error of measurement. Statistical significance was determined using an unpaired t-test (two-tailed). ∗p < 0.05, ∗∗p < 0.01 and denotes comparison with control. D, proteins from the ALA + DFO treated cells were separated by reducing SDS-PAGE and visualized by Coomassie staining (dotted box highlights the stained aggregates that did not enter the gel). The data is representative of three independent experiments.