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Fig. 12 Effect of MRP4 and BSEP siRNA knockdown in ALA+DFO-treated PMH. (A, B) PMH were cultured with scrambled, MRP4, or BSEP siRNA (24 hours) followed by treatment with ALA + DFO (1 mM and 100 μM, respectively) for 12 hours. (A) Immunoblot analyses using antibodies to MRP4 and BSEP. Coomassie staining (CM) and actin blots are included to show equal loading. The MRP4 monomer is indicated by arrowhead. HMW aggregates of MRP4 are highlighted by an arrow. (B) Intracellular and culture medium PP-IX levels following the treatments are shown. (C) Immunoblots of p62 monomer doublet (detected as a doublet due to transfection conditions) and its HMW aggregates in total hepatocyte lysates. Actin blot is included as a loading control. (D) Relative densities of the p62 species from 3 individual experiments were quantified. For panels B and D, data are shown as mean ± SD (N = 3 experiments). Statistical analysis was performed by 2-tailed Student t-test, ∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05.