Knockdown of Heparanase Suppresses Invasion of Human Trophoblasts by Activating p38 MAPK Signaling Pathway

Figure 4

Knockdown of HPSE inhibited trophoblast cell invasion by activating p38 MAPK pathway. (a) Total cell protein including phosphorylated protein was extracted and analyzed by MAPK Pathway Phosphorylation Array C1. 1: p-mTOR. 2: p-p38 MAPK. 3: p-Rsk2. To confirm the results of microarrays, the three phosphorylated proteins and the corresponding total protein were determined in three independent experiments by Western blotting. (b) Western blotting images of p38 MAPK. (c) The gray value ratio of p-p38/t-p38 was elevated in HTR8/SVneo cells transfected with shRNA-HPSE. β-Actin was used as internal control. (d) Representative images of cell invasion after HPSE knockdown in shRNA-HPSE-HTR8 cells, pretreated with pretreated with 1 μM BMS582949, 20 μM SB203580, or 0.5 μM BIRB796 for 2 h. 1: 0.1% DMSO as control. 2: 1 μM BMS582949. 3: 20 μM SB203580. 4: 0.5 μM BIRB796. Magnification: 400x. (e) Quantitative analysis for the number of invasive cells. Data in graph c and graph e were represented as the mean ± SD. The differences among them were compared by one-way ANOVA and Holm-Sidak’s post hoc test. .

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