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. 2019 Mar 9;20(5):1206. doi: 10.3390/ijms20051206

Figure 5.

Figure 5

Inhibition of the Wnt/β-catenin pathway blocks key molecules of lipid metabolism and activation of MARK4 in pig primary trophoblast cells. (A–D) Representative immunoblots and densitometric quantification for p-MARK4 (T214), DKK1 and β-catenin after transfection with Flag-DKK1, sh-DKK1 for 48 h in primary trophoblast cells isolated from pig placentas. Cells were then incubated with 400 μM NEFA or 20 μM Li CL for 24 h (n = 3). (E) Representative images (100×) of β-catenin immunofluorescent staining after transfection with Flag-DKK1, sh-DKK1 for 48 h in pig primary trophoblast cells. Cells were then incubated with 400 μM NEFA or 20 μM Li CL for 24 h (n = 3). (F) Quantification of red fluorescence intensity in (E) relative to control group (n = 3). (G–H) Relative mRNA expression of lipid metabolism-related genes (G) and fatty acid (FA) transporters (H) after transfection with Flag-DKK1, sh-DKK1 for 48 h in primary trophoblast cells. Cells were then treated with 400 μM NEFA or 20 μM Li CL for 24 h (n = 3). Values are expressed as mean ± SEM. * p < 0.05 compared with the control group. Flag-DKK1 group: overexpression of DKK1 group, sh-DKK1 group: knock down of DKK1 group, Control: empty vector (EV) group.