Figure 4. A. Effect of berberine on HIV PI-induced ERK activation in macrophages.

Representative immunoblots from three independent experiments for phospho(p)-ERK and total (T)-ERK from the total cell lysates of mouse J774A.1 cells treated with berberine (BBR, 0.5 µg/ml) and individual HIV PIs (15 µM) for 4 h. ATZV: atazanavir; LOPV: lopinavir; RITV: ritonavir. B: Time course of berberine uptake. Mouse J774A.1 cells were treated with berberine (BBR, 5 µM) for various time periods (0 to 24 h), the concentration of BBR in culture medium and total intracellular amount were determined by HPLC as described in “Methods”. C: Dose-dependence of berberine uptake in macrophages. Cells were treated with various amounts of berberine (BBR, 0.5 to 15 µM) for 1, 8, or 24 h. The intracellular concentrations of BBR were determined as described above and expressed as pmol/mg protein. D. Effect of lopinavir on berberine uptake in macrophages. Mouse J774A.1 cells were treated with berberine (BBR, 5 µM) in the presence or absence of lopinavir (LOPV, 15 µM) for various time periods (0 to 24 h), the concentration of BBR in culture medium and total intracellular amount were determined by HPLC as described above.