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. 2015 Jun 2;5:10729. doi: 10.1038/srep10729

Figure 4. ΔΨm assessed by flow cytometry in VM neurons with blocking LfR.

Figure 4

To block binding of ligands to LfR, VM neurons were treated with Anti-LfR for 1 h at 37 °C. Then, VM neurons were treated with 100 ng/ml apo-Lf or 100 ng/ml holo-Lf for 4 h followed by 100 μmol/L MPP+ for 24 h. (A) Representatives of the fluorometric assay on ΔΨm of different groups. Results are shown as FL1-H (fluorescence 1-histogram), setting the gated region M1 and M2 as a marker to observe the changing levels of fluorescence intensity using CellQuest software. Numbers in the M1 gate of the dot-plots show the percentage of apoptotic cells. The ΔΨm was decreased in MPP+-treated apoptotic cells and the ratio of M1 area was increased. 100 ng/ml apo-Lf or holo-Lf pretreatment significantly restored the ΔΨm reduction induced by MPP+. Blocking LfR significantly attenuated the protective effect of Lf in VM neurons induced by MPP+. X mean = mean channel fluorescence of FL-1; Y mean= the number of living cells. (B) Statistical analysis. Data were presented as mean ± S.E.M. of three independent experiments. Fluorescence values of the control were set to 100%. *P < 0.05, compared with the control; #P < 0.05, compared with MPP+ group; n = 6.