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. 2020 Nov 17;25(22):5375. doi: 10.3390/molecules25225375

Figure 4.

Figure 4

Figure 4

Effect of ethanol extract of Patrinia villosa (Pv-EE) on autophagy. (A–C) Levels of total LC3B, p62, and β-actin proteins were determined in B16F10 cells using the total antibodies for each protein. (D,E) The promoter binding activity of the transcription factor CREB was analyzed using a reporter gene assay. B16F10 cells were transfected with plasmids driving the expression of CREB-Luc (1 μg/mL) and β-gal (as a transfection control). After 24 h, some of the cells were treated with 100 nM α-MSH and lPv-EE (800 μg/mL) or rPv-EE (200 μg/mL) or 3-MA (10 mM) for 48 h. Luciferase activity was measured using a luminometer. (F–H) The levels of melanin secretion and contents in B16F10 cells treated with α-MSH (100 nM) in the presence or absence of lPv-EE (800 μg/mL) or 3-MA (10 mM) or chloroquine (20 μM) or Wortmannin (2 μM) for 48 h were then determined. (I) The phytochemical profile of rPv-EE was analyzed by HPLC using standard compounds (quercetin, luteolin, and kaempferol). ** p < 0.01 compared to the normal group and * p < 0.05 compared to the control group.