Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 1. The expressions of PD-L1 in human preeclampsia (PE) placentas (n = 30) and normal pregnant women (n = 30). (A,B) PD-1 levels were significantly reduced in PE placentas. (C,D) PD-L1 levels were overtly reduced in pre-eclamptic mothers. (E,F) GM-CSF levels were prominently decreased in the PE set. Relative JAK2 (G) and STAT5 (H) protein levels were markedly lessened, while those of p-JAK2 (I) and p-STAT5 (J) were significantly enriched in pre-eclamptic placentas. (K) Western blotting results of related molecules were shown. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques: Western Blot

Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 2. The transfection efficiency in cell lines and the changes in expressions of relevant factors after lentivirus treatment (n = 3, for each group). (A,B) The valid transfection efficiency in the two cell lines was confirmed by the presence of green fluorescence. Scale bars, 100 μm. The mRNA (C,L) and protein (D,M) levels of PD-L1 were significantly reduced after the knockdown of PD-L1 in two cell systems. The mRNA (E,N) and protein (F,O) levels of GM-CSF were notably lower after transfection with siR-PD-L1 in the two trophoblasts. JAK2 and STAT5 protein expressions were induced, while their phosphorylation expressions were inhibited in HTR- 8/SVneo (G–J) and JEG-3 (P–S) cells with the introduction of LV-PD-L1. (K,T) Western blot of PD-L1, GM- CSF, and JAK2/STAT5 molecules was carried out in the two cell lines. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques: Transfection, Fluorescence, Knockdown, Phospho-proteomics, Western Blot

Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 4. Influences of aberrant GM-CSF on PD-L1 and the JAK2/STAT5 pathway (n = 3, for each group). The protein levels (A,H) of PD-L1 were evidently elevated after adding GM-CSF in trophoblasts. GM-CSF protein levels (B,I) were significantly higher by the accumulation of GM-CSF in the two trophoblasts. The levels of JAK2 and STAT5 were overtly lower, while their phosphorylation levels were enhanced exposure to the GM- CSF neutralizing antibody in HTR-8/SVneo (C–F) and JEG-3 (J–M) cells. (G,N) Western blot bands were displayed in the two cell lines. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques: Phospho-proteomics, Western Blot

Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 5. The biological role of GM-CSF in the cell lines (n = 3, for each group). (A,B)Over-regulation of GM- CSF elevated the migratory property of cells. (C,D) GM-CSF stimulation increased cell invasion. Scale bar, 500 μm. (E,F) GM-CSF up-regulation suppressed cell apoptosis. (G–I) Increased GM-CSF promoted the evolvement of cell cycle from the G1 to the S stage in both cells. (J) (K) Cells supplemented with GM-CSF exhibited higher proliferative ability in trophoblasts. ns, no significance. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques:

Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 6. The rescue assay and the impact of the JAK2 inhibitor on regulating GM-CSF (n = 3, for each group). (A,B) The rescue assay of co-transfection of PD-L1 and GM-CSF in two kinds of trophoblasts. The protein levels (C,H) of GM-CSF were elevated with the JAK2 inhibitor. JAK2 (D,I) and STAT5 (E,J) protein levels were not influenced by the JAK2 inhibitor. The levels of p-JAK2 (F,K) and p-STAT5 (G,L) were profoundly inhibited by the addition of JAK2 inhibitor. (M) The bands of western blot in rescue experiments. (N) Western blotting was shown in cells with the JAK2 inhibitor. ns, no significance. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques: Rescue Assay, Cotransfection, Western Blot

Journal: Scientific reports

Article Title: Decreased PD-L1 contributes to preeclampsia by suppressing GM-CSF via the JAK2/STAT5 signal pathway.

doi: 10.1038/s41598-025-87349-1

Figure Lengend Snippet: Fig. 7. The blood pressure, urine protein, and the expressions of related indicators in different groups of pregnant rats (n = 5, for each group). (A,B) LV.PD-L1 reduced the blood pressure of PE-like animals on GD17 and GD19. (C) LV.PD-L1 significantly decreased the urinary protein content on GD19. Fetal weight (D), fetal length (E), and placental weight (F) in different groups. (G) Western blot in rat placental tissues. (H,I) The expressions of PD-1 were evidently reduced after L-NAME treatment. (J,K) The levels of PD-L1 were notably decreased in placentas from PE-like pregnant rats. (L,M) The expressions of GM-CSF were elevated following adding LV.PD-L1. (N–Q) The relative band intensity of the JAK2/STAT5 pathway was compared using quantification. DAPI staining and the green fluorescence representing viral infection were displayed in the L-NAME + PD-L1 NC (R) and L-NAME + LV.PD-L1 (S) sets. Scale bar, 50 μm. Data are analyzed by χ2-test between the two groups.

Article Snippet: In order to test the influence of GM-CSF on the functions of trophoblasts, cell lines were added with 100 ng/ ml recombinant human GM-CSF protein (Peprotech, Rocky Hill, NJ), 10 μg/ml neutralizing GM-CSF antibody, or 10 μg/ml GM-CSF isotype control mAb (Proteintech Group, Rosemont, IL, United States) for 48 h. Similar to another experimental proposal33, recombinant GM-CSF protein was used to treat one set of cell lines, while another set of cell lines did not receive this treatment.

Techniques: Western Blot, Staining, Fluorescence, Infection