Journal: Pharmaceutics

Article Title: Myrrh Oil-Based Nanoemulsion Loaded with Curcumin and Insulin: Development, Characterization, and Evaluation of Enhanced Antibacterial and Diabetic Wound-Healing Activity

doi: 10.3390/pharmaceutics18030369

Figure Lengend Snippet: Effect of topical diabetic wound treatments with various preparations on biomarkers of oxidative stress. ( A ): Nuclear factor erythroid 2-related factor-2 (Nrf-2); ( B ): malondialdehyde (MDA); ( C ): reduced glutathione (GSH). ###: Significantly different compared with the normal control group at p < 0.001, #### at p < 0.0001. *: Significantly different compared with DM + B-NEG group at p < 0.05, ** at p < 0.01, *** at p < 0.001, **** at p < 0.0001. $$$$: Significantly different compared with the CUR-G group at p < 0.0001. @: Significantly different compared with INS-G group at p < 0.05, @@ at p < 0.001.

Article Snippet: An enzyme-linked immunosorbent assay (ELISA) kit was used to measure the concentration of nuclear factor erythroid 2-related factor 2 (Nrf-2, BT LAB, Shanghai, China) in skin tissue homogenate following the manufacturer’s procedures.

Techniques: Control

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 6. Anti-MICA/B 7C6 mAb boosts anti-tumor cytotoxic function of peripheral NK cells from iCCA patients against the HuCCT-1 cell line. A): peripheral NK cell degranulation, evaluated as frequency of CD107a+ NK cells, in iCCA patients (n = 13) and HC (n = 16) in the presence of 7C6 mAb or IgG1-Fc. Parametric paired and unpaired t tests were used to compare data. B): dot plots showing the frequency of CD3-CD56+ CD107a+ NK cells in a HC and a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc. C): the proportion of circulating IFNγ+ NK cells in patients (n = 10) and HC (n = 11) in the presence of 7C6 mAb compared with IgG1-Fc. To compare paired data, the parametric t test and the non-parametric Wilcoxon t test were used. To compare unpaired data, the parametric t test and the non-parametric Mann-Whitney U test were used. D): representative dot plots showing the frequency of CD3-CD56+ IFNγ+ NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: MANN-WHITNEY

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 7. Anti-MICA/B 7C6 mAb boosts anti-tumor cytotoxic function of peripheral NK cells from iCCA patients against patient-derived iCCA cell lines. A): peripheral NK cell degranulation, evaluated as CD107a+NK frequency, in iCCA patients (n = 12) and HC (n = 8) in the presence of 7C6 mAb or IgG1-Fc using patient- derived primary tumor cell lines as targets. Parametric paired and unpaired t tests were used to compare data. B): dot plots showing the frequency of CD3-CD56 + CD107a+ NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc. C): proportion of circulating IFNγ+NK cells in patients (n = 10) and in HC (n = 8) in the presence of 7C6 mAb compared with IgG1-Fc. To compare paired data, we used the parametric t test and the non-parametric Wilcoxon t test. To compare unpaired data, the parametric t test and the non-parametric Mann-Whitney U test were used. D): representative dot plots showing the frequency of CD3-CD56+ IFNγ +NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Derivative Assay, MANN-WHITNEY

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 8. 7C6 mAb enhances the anti-tumor effect of liver- and tumor-infiltrating NK cells in iCCA patients. A): Frequency of degranulating CD107a+NK cells in LIL (n = 13) and TIL (n = 10) of iCCA patients in the presence of anti-MICA/B 7C6 mAb or IgG1-Fc using autologous tumor-derived cell lines as targets. Parametric paired and unpaired t tests were used to compare data. B): representative dot plots showing the frequency of CD3-CD56+ CD107a+ LIL- and TIL-NK cells in the presence of 7C6 mAb or IgG1-Fc. C): proportion of IFNγ+ NK cells in LIL (n = 10) and TIL (n = 8) of iCCA patients in the presence of 7C6 mAb compared with IgG1-Fc using autologous tumor-derived cell lines as targets. The parametric t test and non-parametric Wilcoxon t test were used to compare paired data. The parametric t test was used to compare unpaired data.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Derivative Assay

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 9. Cytotoxicity assay of HC PBMC, patient PBMC, LIL and TIL cells. A, B): Frequency of CFSE+LIVE/DEAD (LD)+ HuCCT-1 cell line targets when HC PBMC (n = 5), patient PBMC (n = 10), LIL (n = 8) and TIL (n = 5) were used as effector cells in the presence of 7C6 mAb and isotype control (IgG1). The parametric paired t tests were used to compare data. C, D): Frequency of CFSE+LD+ patient-derived cell line targets when HC PBMC (n = 5), patient PBMC (n = 8), LIL (n = 8) and TIL (n = 4) were used as effectors in the presence of 7C6 and isotype control. The parametric paired t test was used to compare data in panel C. The non-parametric Wilcoxon t test was used to compare paired data in panel D. Target cell death was determined as frequency of CFSE+LD+ cells.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Cytotoxicity Assay, Control, Derivative Assay

Journal: Cancer Research

Article Title: BRAF Inhibitors Reprogram Cancer-Associated Fibroblasts to Drive Matrix Remodeling and Therapeutic Escape in Melanoma

doi: 10.1158/0008-5472.can-21-0614

Figure Lengend Snippet: Figure 1. BRAFi drives increased nuclear b-catenin in CAFs. A–D, Human melanoma sections [collected from eight patients with melanoma before (A) and after (B) BRAFi/ MEKi therapy] were coimmunostained for aSMA (green) and b-catenin expression (red). Nuclei were stained blue with DAPI. C and D are close-up pictures of the area circled by white boxes in A and B, respectively. Yellow triangles in D indicate aSMAþ fibroblasts expressing nuclear b-catenin. Scale bar, 100 mm. E, The scattered plot represents the numbers of aSMAþ cells per mm2 counted in pre- and posttreated melanoma samples. F, Percentage of aSMAþ cells expressing nuclear b-catenin in pre- and posttreated melanoma samples. Each data point represents the quantification result of one picture. At least five pictures were taken per tumor section. n ≥40. G–L, Immunostaining of b-catenin in M50 treated with DMSO or indicated inhibitors for 72 hours. M, Quantification and comparison of relative fluorescence intensities of nuclear b-catenin in M50 treated with DMSO or indicated BRAFi/MEKi for 72 hours. The fluorescence intensity of nuclear b-catenin in DMSO-treated M50 was used as the control and set at one for comparison. Each data point represents the relative fluorescent intensities of nuclear b-catenin in one treated cell. n ¼ 30. N, Nuclear and cytoplasmic fractions of M50 treated with DMSO or different concentrations of PLX4032. Lamin B was used as a nuclear loading control and tubulin as a cytoplasmic loading control. O and P, Quantification of fluorescence intensities of nuclear and cytoplasmic b-catenin Western blot bands. n ¼ 3. For all graphs, data are represented as mean SEM. , P ≤0.05; , P ≤0.001; ns, not significant.

Article Snippet: Human melanoma cell lines, including 1205Lu (1205 Lu-01–0001, RRID:CVCL_5239), WM278 (WM278–01–0001, RRID:CVCL_6473), and WM1366 (WM1366–01–0001, RRID:CVCL_6789), were purchased from Rockland Immunochemicals.

Techniques: Expressing, Staining, Immunostaining, Comparison, Control, Western Blot

Journal: Cancer Research

Article Title: BRAF Inhibitors Reprogram Cancer-Associated Fibroblasts to Drive Matrix Remodeling and Therapeutic Escape in Melanoma

doi: 10.1158/0008-5472.can-21-0614

Figure Lengend Snippet: Figure 5. POSTN is a direct downstream effector of nuclear b-catenin signaling in CAFs. A, Fibroblasts and POSTN expression in human melanomas pre- and post-BRAFi/MEKi treatment were visualized by coimmunostaining using an anti-TE7 antibody (green) and an anti-POSTN antibody (red). Scale bar, 100 mm. B, The scatter plot represents the percentage of TE7þ cells expressing POSTN in pre- and posttreated melanoma samples. Each data point represents the percentage of TE7þ fibroblasts that are POSTNþ in each 40field. n ≥40. C, Western blotting of POSTN expression levels in indicated melanoma cells and CAFs. Chart shows fluorescence intensities of POSTN bands. n ¼ 3. D, Graph shows qPCR data of POSTN expression in A375, GFP/M50, and bcat-GFP/M50 treated with DMSO or PLX4032. n ¼ 4. E, Illustration of the promoter region of POSTN that contains a potential LEF/TCF–binding site. F, Bar graph shows signals obtained from chromatin immunoprecipitation normalized by signals obtained from the input sample. Axin 2 was used as a positive control. Three different primer sets targeting the TCF/LEF–binding sites in the POSTN promoter region (POSTN1, POSTN2, and POSTN3) and three negative control primers targeting the 30UTR, exon 6, and the 50UTR were used. G, Western blotting of POSTN expression in M50 transfected with scramble siRNA or POSTN siRNA. Graph shows fluorescence intensities of POSTN bands. n ¼ 3. H and I, Graph shows A375 numbers in cocultured spheroids of indicated groups under PLX4032 treatment (H) or PLX4032 and GDC0973 treatment (I). n ¼ 3. J and K, Graph shows A375 numbers in cocultured spheroids of indicated groups with or without recombinant POSTN under PLX4032 treatment (J) or PLX4032 and GDC0973 treatment (K). n ¼ 3. For all graphs, the data are shown as mean SEM. , P ≤0.05; , P ≤0.01; , P ≤0.001. ns, not significant.

Article Snippet: Human melanoma cell lines, including 1205Lu (1205 Lu-01–0001, RRID:CVCL_5239), WM278 (WM278–01–0001, RRID:CVCL_6473), and WM1366 (WM1366–01–0001, RRID:CVCL_6789), were purchased from Rockland Immunochemicals.

Techniques: Expressing, Western Blot, Binding Assay, Chromatin Immunoprecipitation, Positive Control, Negative Control, Transfection, Recombinant

Journal: FEBS Open Bio

Article Title: High glucose stimulates expression of aldosterone synthase ( CYP 11B2 ) and secretion of aldosterone in human adrenal cells

doi: 10.1002/2211-5463.12277

Figure Lengend Snippet: Effects of ARB s (A) and CCB s (B) on the high glucose‐induced CYP 11B2 transactivation. In (A), CYP 11B2 ‐H295R cells were incubated with either 100 mg·dL −1 d ‐glu (control), 450 mg·dL −1 d ‐glu ( d ‐glu), 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 losartan ( d ‐glu + Los), 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 valsartan ( d ‐glu + Val), 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 olmesartan ( d ‐glu + Olm), or 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 candesartan ( d ‐glu + Can) for 48 h. Data represent mean ± SEM ( n = 3), percentage of control. * P < 0.01, vs. control. In (B), CYP 11B2 ‐H295R cells were incubated with either 100 mg·dL −1 d ‐glu, 450 mg·dL −1 d ‐glu, 450 mg·dL −1 d ‐glu plus 0.01 μmol·L −1 amlodipine, 450 mg·dL −1 d ‐glu plus 0.1 μmol·L −1 amlodipine, 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 amlodipine, 450 mg·dL −1 d ‐glu plus 0.01 μmol·L −1 benidipine, 450 mg·dL −1 d ‐glu plus 0.1 μmol·L −1 benidipine, 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 benidipine, 450 mg·dL −1 d ‐glu plus 0.01 μmol·L −1 efonidipine, 450 mg·dL −1 d ‐glu plus 0.1 μmol·L −1 efonidipine, 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 efonidipine, 450 mg·dL −1 d ‐glu plus 0.01 μmol·L −1 nifedipine, 450 mg·dL −1 d ‐glu plus 0.1 μmol·L −1 nifedipine, or 450 mg·dL −1 d ‐glu plus 1 μmol·L −1 nifedipine for 96 h. Data represent mean ± SEM ( n = 4), percentage of 100 mg·dL −1 d ‐glu. * P < 0.01, vs. 100 mg·dL −1 d ‐glu. ** P < 0.05, vs. 450 mg·dL −1 d ‐glu. *** P < 0.01, vs. 450 mg·dL −1 d ‐glu.

Article Snippet: Olmesartan (olmesartan medoxomil) was purchased from Toronto Research Chemicals (North York, Canada).

Techniques: Incubation, Control

Journal: Nature Communications

Article Title: Non-canonical pathway for Rb inactivation and external signaling coordinate cell-cycle entry without CDK4/6 activity

doi: 10.1038/s41467-023-43716-y

Figure Lengend Snippet: a Density scatterplot of Hoechst and EdU staining in MP41 cells treated with palbociclib (1 µM) for the indicated time ( n = 1000 cells/condition). b Immunoblot showing Rb and GAPDH expression in MP41 cells after palbociclib (1 µM) treatment. c Immunoblot showing p-ERK, t-ERK, c-Myc, and GAPDH expression in H1373, WM983B, WM989, and MP41 cells treated with DMSO or the indicated drug for 24 h (sotorasib, 1 µM; vemurafenib, 1 µM; trametinib, 10 nM). d EU levels 24 h after treatment with the indicated drug. Data are shown as mean ± SD ( n = 3 biological replicates). Asterisks indicate significant differences in the two-tailed unpaired t -test (* p ≤ 0.05; ** p ≤ 0.001). e Representative histograms of EdU levels in H1373 cells showing classification of S-phase cells. 24 h after treatment with the indicated drug (palbociclib, 1 µM; sotorasib, 1 µM), cells were incubated with EdU (10 µM) for 48 h prior to fixation ( n > 3000 cells/condition). f Percentage of S-phase cells in H1373, WM989, and MP41 cells. Data are shown as mean ± SD ( n = 3 biological replicates). Asterisks indicate significant differences in the one-way ANOVA test (* p ≤ 0.05; ** p ≤ 0.001; *** p ≤ 0.0001). g Immunoblot showing Rb, c-Myc, and GAPDH expression in H1373, WM989, and MP41 cells 72 h after treatment with the indicated drug.

Article Snippet: WM989 (Rockland Immunochemicals, #WM989-01-0001) and WM983B cells (Rockland Immunochemicals, #WM983B-01-0001) were cultured in a mixture of 80% MCDB153 (Sigma, #M7403) and 20% Leibovitz’s L15 media (Gibco, #21083027), supplemented with 2% FBS and 1.68 mM CaCl 2 .

Techniques: Staining, Western Blot, Expressing, Two Tailed Test, Incubation

Journal: iScience

Article Title: A pharmacological rat model of recurrent pelvic pain exhibiting hyperalgesia and depression-like behaviors

doi: 10.1016/j.isci.2026.115059

Figure Lengend Snippet: Persistent hyperalgesia in the RPP model (A and B) Paw withdrawal latency (A) and 50% paw withdrawal threshold (B) of each rat ( n = 3–6). (C) Representative histological images of DRG from T10-S4 by H&E staining. Arrow, Schwann cell; ∗, nuclei; #, cytoplasm. Scale bars, 50 μm and 25 μm. (D and E) Representative IHC images (D) and quantification (E) of BDNF in the DRG from T10-S4. Brown DAB staining indicates BDNF immunopositivity. The staining intensity was semi-quantitatively scored as follows: (1) weak, (2) moderate, and (3) intense. Scale bars, 50 μm. ( n = 3 biological replicates per group). (F) Serum BDNF level on days 12 and 24 of RPP modeling ( n = 3). Data are presented as the mean ± SD; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, two-way ANOVA (A and B) or one-way ANOVA (E and F).

Article Snippet: Rat BDNF ELISA Kit , Elabscience , Cat# E-EL-R1235.

Techniques: Staining

Journal: iScience

Article Title: A pharmacological rat model of recurrent pelvic pain exhibiting hyperalgesia and depression-like behaviors

doi: 10.1016/j.isci.2026.115059

Figure Lengend Snippet: Comorbid depression-like behavior in the RPP model (A and B) Serum BDNF (A) and 5-HT (B) levels in the RPP model ( n = 6). (C and D) Sucrose preference ratio (C) and immobility time (D) in the RPP model ( n = 8). Data are presented as the mean ± SD; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, paired t test (A and B) or two-way ANOVA (C and D).

Article Snippet: Rat BDNF ELISA Kit , Elabscience , Cat# E-EL-R1235.

Techniques:

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: BMP6 as a therapeutic target for preeclampsia: enhancing trophoblast invasion and vascular mimicry

doi: 10.1007/s00018-025-06040-w

Figure Lengend Snippet: BMP6 signaling enhancement compensates for shallow trophoblast invasion in PE. An examination of clinical samples from PE patients and their gestational age-matched controls revealed that BMP6 was upregulated in preeclamptic placentas. BMP6 enhances trophoblast invasion via ID1-mediated upregulation of SERPINE2 and PlGF. Moreover, BMP6 also intensifies trophoblast vascular mimicry through the ID1-mediated upregulation of PlGF. Both canonical (p-SMAD1/5/9) and noncanonical (p-SMAD2/3) pathways participate in BMP6-induced ID1 expression. Our findings indicate that the increase in BMP6 signaling during late gestation could serve as a compensatory response to shallow trophoblast invasion in PE, which in light of BMP6 and its downstream targets could serve as diagnostic markers and therapeutic target applications in the clinical management of PE. This schematic diagram was created with Biorender.com

Article Snippet: SMAD1 (D59D7) rabbit monoclonal antibody (#6944), phospho-SMAD1/5/9 (D5B10) rabbit monoclonal antibody (#13820), SMAD2 (D43B4) rabbit monoclonal antibody (#5339), phospho-SMAD2 (E8F3R) rabbit monoclonal antibody (#18338), SMAD3 (C67H9) rabbit monoclonal antibody (#9523), phospho-SMAD3 (C25A9) rabbit monoclonal antibody (# 9520), SMAD4 (D3R4N) rabbit monoclonal antibody (#46535), horseradish peroxidase (HRP)-linked anti-mouse IgG (#7076), and anti-rabbit IgG (#7074) were purchased from Cell Signaling Technology (Danvers, MA, USA); the mouse monoclonal antibody HLA-G (#11–499-C100) was purchased from EXBIO (Vestec, Czech Republic); CK7-specific rabbit polyclonal antibody (#17513-1-AP), BMP6 rabbit polyclonal antibody (#55421-1-AP), SERPINE2 rabbit polyclonal antibody (#11303-1-AP), and α-Tubulin mouse monoclonal antibody (#66031-1-Ig) were purchased from Proteintech (Wuhan, China); ID1 mouse monoclonal antibody (#sc-133104) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA); CD34 rabbit monoclonal antibody (#A19015) and α-SMA rabbit monoclonal antibody (#A2235) were obtained from ABclonal (Wuhan, China); Goat anti-Rabbit IgG secondary antibody, Alexa Fluor 594 (#A-11012) and goat anti-Mouse IgG secondary antibody, Alexa Fluor 488 (#A-11011) were purchased from Thermo Fisher (NY, USA).

Techniques: Expressing, Diagnostic Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: BMP6 as a therapeutic target for preeclampsia: enhancing trophoblast invasion and vascular mimicry

doi: 10.1007/s00018-025-06040-w

Figure Lengend Snippet: ID1 mediates BMP6-induced SERPINE2 and PlGF upregulation in human trophoblasts. A-C , BMP6 upregulates SERPINE2 protein levels in trophoblasts. A , HTR8/SVneo cells were treated with different concentrations (0, 6.25, 12.5, 25, 50, or 100 ng/mL) of BMP6, and the SERPINE2 protein levels after 24 h of treatment were examined by Western blot analysis. The upper panel shows a representative Western blot image, and the lower panel shows the summarized quantitative results. B , SERPINE2 protein levels in HTR8/SVneo cells after treatment with vehicle (Ctrl) or 50 ng/mL BMP6 for different durations (24, 48, and 72 h). The upper panel shows a representative Western blot image, and the lower panel shows the summarized quantitative results. C , SERPINE2 protein levels in human primary EVTs. The left panel shows a representative Western blot image, and the right panel shows the summarized quantitative results. D-E , BMP6 promotes PlGF accumulation in the conditioned medium of trophoblasts. D , HTR8/SVneo cells were treated with or without 50 ng/mL BMP6 for 24–48 h. PlGF accumulation in conditioned medium was measured using ELISA. E , PlGF accumulation in conditioned medium was assayed by ELISA 48 h after BMP6 treatment in primary EVTs. F-J , ID1 mediates BMP6-induced SERPINE2 and PlGF upregulation in human trophoblasts. HTR8/SVneo cells were transfected for 48 h with 20 nM control non-targeting siRNA (si-Ctrl) or siRNA targeting ID1 (si- ID1 ) before treatment with or without 50 ng/mL BMP6. F , ID1 mRNA levels were examined by qPCR 6 h after BMP6 (50 ng/mL) treatment in HTR8/SVneo cells, with GAPDH used as the reference gene. G and H , SERPINE2 and ID1 protein levels in HTR8/SVneo cells ( G ) and human primary EVTs ( H ) after transfection with siRNA targeting ID1 , followed by treatment with or without BMP6 for 24 h, as assessed by Western blot. The left panel shows a representative Western blot image, and the right panel shows the summarized quantitative results. I , PGF mRNA levels were examined by qPCR 6 h after BMP6 treatment in HTR8/SVneo cells, with GAPDH used as the reference gene. J , PlGF accumulation in conditioned medium was assayed by ELISA 24 h after BMP6 treatment in HTR8/SVneo cells. K , SMAD4 mediates BMP6-induced upregulation of PlGF in human trophoblasts. HTR8/SVneo cells were transfected for 48 h with 20 nM control non-targeting siRNA (si-Ctrl) or siRNA targeting SMAD4 (si- SMAD4 ) before treatment with or without 50 ng/mL BMP6. PlGF accumulation in conditioned medium was assayed by ELISA 24 h after BMP6 treatment in HTR8/SVneo cells. The quantitative results are expressed as the means ± SEMs of at least three independent experiments. One-way ANOVA was used for analyses in A , and two-way ANOVA was used for comparisons in B-K . Groups without common letters are significantly different from each other ( P < 0.05). BMP6, bone morphogenetic protein 6; SERPINE2, serpin family E member 2; EVT, extravillous cytotrophoblast; Ctrl, control; PlGF, placental growth factor; ID1, inhibitor of DNA-binding 1

Article Snippet: SMAD1 (D59D7) rabbit monoclonal antibody (#6944), phospho-SMAD1/5/9 (D5B10) rabbit monoclonal antibody (#13820), SMAD2 (D43B4) rabbit monoclonal antibody (#5339), phospho-SMAD2 (E8F3R) rabbit monoclonal antibody (#18338), SMAD3 (C67H9) rabbit monoclonal antibody (#9523), phospho-SMAD3 (C25A9) rabbit monoclonal antibody (# 9520), SMAD4 (D3R4N) rabbit monoclonal antibody (#46535), horseradish peroxidase (HRP)-linked anti-mouse IgG (#7076), and anti-rabbit IgG (#7074) were purchased from Cell Signaling Technology (Danvers, MA, USA); the mouse monoclonal antibody HLA-G (#11–499-C100) was purchased from EXBIO (Vestec, Czech Republic); CK7-specific rabbit polyclonal antibody (#17513-1-AP), BMP6 rabbit polyclonal antibody (#55421-1-AP), SERPINE2 rabbit polyclonal antibody (#11303-1-AP), and α-Tubulin mouse monoclonal antibody (#66031-1-Ig) were purchased from Proteintech (Wuhan, China); ID1 mouse monoclonal antibody (#sc-133104) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA); CD34 rabbit monoclonal antibody (#A19015) and α-SMA rabbit monoclonal antibody (#A2235) were obtained from ABclonal (Wuhan, China); Goat anti-Rabbit IgG secondary antibody, Alexa Fluor 594 (#A-11012) and goat anti-Mouse IgG secondary antibody, Alexa Fluor 488 (#A-11011) were purchased from Thermo Fisher (NY, USA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Control, Binding Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: BMP6 as a therapeutic target for preeclampsia: enhancing trophoblast invasion and vascular mimicry

doi: 10.1007/s00018-025-06040-w

Figure Lengend Snippet: Both SERPINE2 and PlGF mediate BMP6-induced trophoblast invasion. A-J , HTR8/SVneo cells or primary EVTs were transfected for 48 h with 20 nM control nontargeting siRNA (si-Ctrl), 20 nM siRNA targeting SERPINE2 (si- SERPINE2 ) or PGF (si- PGF ) before treatment with or without 50 ng/mL BMP6 for 24 h. A and B , The protein levels of SERPINE2 in HTR8/SVneo cells ( A ) and human primary EVTs ( B ) after 24 h of BMP6 treatment. The upper panel shows a representative Western blot image, and the lower panel shows the summarized quantitative results. C , PlGF accumulation in conditioned medium with or without BMP6 treatment for 24 h was assayed by ELISA in HTR8/SVneo cells. D , PGF mRNA levels in human primary EVTs with or without BMP6 treatment for 6 h were examined by RT‒qPCR, with GAPDH as the reference gene. E - H , Transwell assays were employed to examine the invasiveness of HTR8/SVneo cells ( E and G ) and primary EVTs ( F and H ) with or without BMP6 treatment for 36 h. I and J , Endothelial-like tube formation assays were used to assess vascular mimicry of HTR8/SVneo cells with or without BMP6 treatment for 12 h. Representative images from the endothelial-like tube formation assay are displayed in the above panel; the summarized quantitative results are displayed in the lower panel. Scale bar, 100 μm. The quantitative results are expressed as the means ± SEMs of at least three independent experiments. Two-way ANOVA was used for data comparison. Groups without common letters are significantly different from each other ( P < 0.05). BMP6, bone morphogenetic protein 6; SERPINE2, serpin family E member 2; PGF, placental growth factor; Ctrl, control; EVT, extravillous cytotrophoblast

Article Snippet: SMAD1 (D59D7) rabbit monoclonal antibody (#6944), phospho-SMAD1/5/9 (D5B10) rabbit monoclonal antibody (#13820), SMAD2 (D43B4) rabbit monoclonal antibody (#5339), phospho-SMAD2 (E8F3R) rabbit monoclonal antibody (#18338), SMAD3 (C67H9) rabbit monoclonal antibody (#9523), phospho-SMAD3 (C25A9) rabbit monoclonal antibody (# 9520), SMAD4 (D3R4N) rabbit monoclonal antibody (#46535), horseradish peroxidase (HRP)-linked anti-mouse IgG (#7076), and anti-rabbit IgG (#7074) were purchased from Cell Signaling Technology (Danvers, MA, USA); the mouse monoclonal antibody HLA-G (#11–499-C100) was purchased from EXBIO (Vestec, Czech Republic); CK7-specific rabbit polyclonal antibody (#17513-1-AP), BMP6 rabbit polyclonal antibody (#55421-1-AP), SERPINE2 rabbit polyclonal antibody (#11303-1-AP), and α-Tubulin mouse monoclonal antibody (#66031-1-Ig) were purchased from Proteintech (Wuhan, China); ID1 mouse monoclonal antibody (#sc-133104) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA); CD34 rabbit monoclonal antibody (#A19015) and α-SMA rabbit monoclonal antibody (#A2235) were obtained from ABclonal (Wuhan, China); Goat anti-Rabbit IgG secondary antibody, Alexa Fluor 594 (#A-11012) and goat anti-Mouse IgG secondary antibody, Alexa Fluor 488 (#A-11011) were purchased from Thermo Fisher (NY, USA).

Techniques: Transfection, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Tube Formation Assay, Comparison

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: BMP6 as a therapeutic target for preeclampsia: enhancing trophoblast invasion and vascular mimicry

doi: 10.1007/s00018-025-06040-w

Figure Lengend Snippet: BMP6 is elevated in patients with PE and in PE model rats. A-C , BMP6 is elevated in patients with PE. A , RT‒qPCR analysis of BMP6 mRNA expression levels in the placentas of control women ( n = 10) and PE patients ( n = 10), with GAPDH as the reference gene. B , Western blot analysis of BMP6 protein expression levels in the placentas of control women ( n = 4) and PE patients ( n = 4). C , Spearman correlation analysis between the placental BMP6 mRNA levels and the value of log10 (SBP) of the corresponding patients. The gray area represents the 95% CI. Each dot represents one sample. D , The animal experimental protocol. E and F , BMP6 is elevated in the plasma of PE model rats. Rat plasma levels of BMP6 ( E ) and PlGF ( F ) in the Ad Fc + PBS group ( n = 4) and Ad Flt1 + PBS group ( n = 4). G-M , BMP6 is elevated in the placenta of PE model rats at G13. RNA-seq analysis of rat placentas at G13 in the Ad Fc + PBS group ( n = 3) and Ad Flt1 + PBS group ( n = 3). G , Heatmap depicting DEGs in the two groups. H , Dot plots of significantly enriched GO terms; the dot size represents the number of DEGs associated with a particular GO term. I , KEGG hierarchical network plot of pathways. J , GSEA-KEGG Ridge plot of pathways. K , GSEA plots of cytokine-cytokine receptor interaction pathway. L , Volcano plot of RNA-seq data showing DEGs between the Ad Fc + PBS group and the Ad Flt1 + PBS group. M , Circos graph displaying the coexpression networks of five genes in rat placenta samples. Each sector of the circle represents one gene, and its width indicates the total amount of co-occurrence that connects one gene to the other. The width of each link represents the total number of coexpressed genes among the linked genes. Student’s t-test was used for comparisons between two groups in A , B , E , and F . Groups without common letters are significantly different from each other ( P < 0.05). SD, Sprague–Dawley; BMP6, bone morphogenetic protein 6; PlGF, placental growth factor; PBS, phosphate-buffered saline; Ad Flt1, adenovirus expressing fms-like tyrosine kinase-1; Ad Fc, adenovirus-expressing control IgG2a Fc fragment; FC, fold change; Serpine2, serpin family E member 2; Id1, inhibitor of DNA-binding 1

Article Snippet: SMAD1 (D59D7) rabbit monoclonal antibody (#6944), phospho-SMAD1/5/9 (D5B10) rabbit monoclonal antibody (#13820), SMAD2 (D43B4) rabbit monoclonal antibody (#5339), phospho-SMAD2 (E8F3R) rabbit monoclonal antibody (#18338), SMAD3 (C67H9) rabbit monoclonal antibody (#9523), phospho-SMAD3 (C25A9) rabbit monoclonal antibody (# 9520), SMAD4 (D3R4N) rabbit monoclonal antibody (#46535), horseradish peroxidase (HRP)-linked anti-mouse IgG (#7076), and anti-rabbit IgG (#7074) were purchased from Cell Signaling Technology (Danvers, MA, USA); the mouse monoclonal antibody HLA-G (#11–499-C100) was purchased from EXBIO (Vestec, Czech Republic); CK7-specific rabbit polyclonal antibody (#17513-1-AP), BMP6 rabbit polyclonal antibody (#55421-1-AP), SERPINE2 rabbit polyclonal antibody (#11303-1-AP), and α-Tubulin mouse monoclonal antibody (#66031-1-Ig) were purchased from Proteintech (Wuhan, China); ID1 mouse monoclonal antibody (#sc-133104) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA); CD34 rabbit monoclonal antibody (#A19015) and α-SMA rabbit monoclonal antibody (#A2235) were obtained from ABclonal (Wuhan, China); Goat anti-Rabbit IgG secondary antibody, Alexa Fluor 594 (#A-11012) and goat anti-Mouse IgG secondary antibody, Alexa Fluor 488 (#A-11011) were purchased from Thermo Fisher (NY, USA).

Techniques: Expressing, Control, Western Blot, Clinical Proteomics, RNA Sequencing, Saline, Binding Assay