Figures & data
Figure 1. Downregulation of PTPRO promoted the proliferation of trophoblast cells. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO-1/2 or siRNA-NC. (a) RT-qPCR was employed to detect the mRNA level of PTPRO in HTR-8/SVneo cells. (b) Western blot analysis was employed to detect the protein level of PTPRO in HTR-8/SVneo cells. (c) RT-qPCR was employed to detect the mRNA level of PTPRO in JEG-3 cells. (d) Western blot analysis was employed to detect the protein level of PTPRO in JEG-3 cells. (e) CCK-8 assay was employed to assess the proliferation of HTR-8/SVneo cells. (f) CCK-8 assay was employed to assess the proliferation of JEG-3 cells. (g) Western blot analysis was employed to detect the protein level of Ki67 in HTR-8/SVneo cells. (h) Western blot analysis was employed to detect the protein level of Ki67 in JEG-3 cells. **p < 0.01, ***p < 0.001
Figure 2. Downregulation of PTPRO promoted cell cycle progression. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or siRNA-NC. (a, b) Flow cytometry was adopted to evaluate the cell cycle distribution of HTR-8/SVneo cells. (c, d) Flow cytometry was adopted to evaluate the cell cycle distribution of JEG-3 cells. ***p < 0.001
Figure 3. Downregulation of PTPRO facilitated the invasion of trophoblast cells. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or siRNA-NC. (a, b) Transwell assay was performed to evaluate the invasion of HTR-8/SVneo cells. (c, d) Transwell assay was performed to evaluate the invasion of JEG-3 cells. (e) RT-qPCR was employed to detect the mRNA levels of MMP2 and MMP9 in HTR-8/SVneo cells. (f) Western blot analysis was employed to detect the protein levels of MMP2 and MMP9 in HTR-8/SVneo cells. (g) RT-qPCR was employed to detect the mRNA levels of MMP2 and MMP9 in JEG-3 cells. (h) Western blot analysis was employed to detect the protein levels of MMP2 and MMP9 in JEG-3 cells. **p < 0.01, ***p < 0.001
Figure 4. Downregulation of PTPRO induced a stronger in vitro angiogenesis. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or siRNA-NC. HUVECs were incubated with the conditioned media (CM) of HTR-8/SVneo or JEG-3 cells. (a-c) The angiogenic activity of HUVECs incubated with the conditioned media (CM) of HTR-8/SVneo cells was tested using an in vitro tube formation assay. (d-f) The angiogenic activity of HUVECs incubated with the conditioned media (CM) of JEG-3 cells was tested using an in vitro tube formation assay. (g) Western blot analysis was employed to detect the protein levels of VEGF, ET-1 and sFlt-1 in HUVECs incubated with the conditioned media (CM) of HTR-8/SVneo cells. (h) Western blot analysis was employed to detect the protein levels of VEGF, ET-1 and sFlt-1 in HUVECs incubated with the conditioned media (CM) of JEG-3 cells. **p < 0.01, ***p < 0.001
Figure 5. PTPRO interacted with ERp44 to participate in PE progression. (a-d) Co-IP assays were carried out to determine the interaction between PTPRO and ERp44. (e, f) HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or siRNA-NC. Western blot analysis was employed to detect the protein level of ERp44 in HTR-8/SVneo and JEG-3 cells. ***p < 0.001
Figure 6. Downregulation of PTPRO promoted the proliferation of trophoblast cells by suppressing ERp44 expression. (a, b) HTR-8/SVneo and JEG-3 cells were transfected with Ov-ERp44 or Ov-NC. RT-qPCR was employed to detect the mRNA level of ERp44 in HTR-8/SVneo and JEG-3 cells. (c, d) HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or co-transfected with siRNA-PTPRO and Ov-ERp44. CCK-8 assay was employed to assess the proliferation of HTR-8/SVneo and JEG-3 cells. (e, f) HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or co-transfected with siRNA-PTPRO and Ov-ERp44. Western blot analysis was employed to detect the protein level of Ki67 in HTR-8/SVneo and JEG-3 cells. *p < 0.05, **p < 0.01, ***p < 0.001; #p < 0.05, ##p < 0.01, ###p < 0.001
Figure 7. Downregulation of PTPRO promoted cell cycle progression by suppressing ERp44 expression. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or co-transfected with siRNA-PTPRO and Ov-ERp44. (a, b) Flow cytometry was adopted to evaluate the cell cycle distribution of HTR-8/SVneo cells. (c, d) Flow cytometry was adopted to evaluate the cell cycle distribution of JEG-3 cells. *p < 0.05, **p < 0.01, ***p < 0.001
Figure 8. Downregulation of PTPRO facilitated the invasion of trophoblast cells by suppressing ERp44 expression. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or co-transfected with siRNA-PTPRO and Ov-ERp44. (a, b) Transwell assay was performed to evaluate the invasion of HTR-8/SVneo cells. (c, d) Transwell assay was performed to evaluate the invasion of JEG-3 cells. (e) RT-qPCR was employed to detect the mRNA levels of MMP2 and MMP9 in HTR-8/SVneo cells. (f) Western blot analysis was employed to detect the protein levels of MMP2 and MMP9 in HTR-8/SVneo cells. (g) RT-qPCR was employed to detect the mRNA levels of MMP2 and MMP9 in JEG-3 cells. (h) Western blot analysis was employed to detect the protein levels of MMP2 and MMP9 in JEG-3 cells. * p < 0.05, **p < 0.01, ***p < 0.001
Figure 9. Downregulation of PTPRO induced a stronger in vitro angiogenesis by suppressing ERp44 expression. HTR-8/SVneo and JEG-3 cells were transfected with siRNA-PTPRO or co-transfected with siRNA-PTPRO and Ov-ERp44. HUVECs were incubated with the conditioned media (CM) of HTR-8/SVneo or JEG-3 cells. (a-c) The angiogenic activity of HUVECs incubated with the conditioned media (CM) of HTR-8/SVneo cells was tested using an in vitro tube formation assay. (d-f) The angiogenic activity of HUVECs incubated with the conditioned media (CM) of JEG-3 cells was tested using an in vitro tube formation assay. (g) Western blot analysis was employed to detect the protein levels of VEGF, ET-1 and sFlt-1 in HUVECs incubated with the conditioned media (CM) of HTR-8/SVneo cells. (h) Western blot analysis was employed to detect the protein levels of VEGF, ET-1 and sFlt-1 in HUVECs incubated with the conditioned media (CM) of JEG-3 cells. **p < 0.01, ***p < 0.001
Availability of data and materials
The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.