Journal: Scientific Reports

Article Title: BMP2 expression in oral squamous cell carcinoma and its effects on SCC9 cell biological behavior

doi: 10.1038/s41598-025-96274-2

Figure Lengend Snippet: Protein expression profiles were analyzed through immunoblotting across experimental groups. Notably, GAPDH, Smad1/5, and p38 protein levels remained unaltered regardless of BMP2 expression status in SCC9 cells. BMP2 overexpression induced significant phosphorylation enhancement of Smad1/5 (52% increase, p = 0.017) and p38 (33% increase, p = 0.020). Conversely, BMP2 knockdown substantially attenuated Smad1/5 phosphorylation (76% reduction, p = 0.009) and p38 phosphorylation (34% reduction, p = 0.045), with asterisks denoting statistical significance. * p < 0.05, ** p < 0.01.

Article Snippet: Experiments in this study were conducted with the following components: fetal bovine serum (Life-ilab, China); Cell Counting Kit-8 (CCK8) kit (ZomanBio, China); Transwell chambers (Corning, USA); Radio Immunoprecipitation Assay Lysis buffer, anti-mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, and anti-rabbit Smad1/5 polyclonal antibody (Boster Bio, China); anti-rabbit p-Smad1/5 (Bioss, China); anti-rabbit BMP2 polyclonal antibody, anti-rabbit p38 polyclonal antibody, and anti-rabbit p-p38 polyclonal antibody (Wanlei Biology, Liaoning); BMP2 small interfering RNA (si-BMP2), control small interfering RNA (si-NC), BMP2 -OE plasmid, control plasmid, and Transfect-Mate transfection reagent (GenePharma, China); and Quantitative Rea-ltime-PCR (qPCR) primers (Thermo Fisher, USA).

Techniques: Expressing, Western Blot, Over Expression, Knockdown

Journal: Scientific Reports

Article Title: BMP2 expression in oral squamous cell carcinoma and its effects on SCC9 cell biological behavior

doi: 10.1038/s41598-025-96274-2

Figure Lengend Snippet: The mRNA expression levels of Smad1/4/5 and p38 in SCC9 cells were quantitatively analyzed using qPCR under different BMP2 modulation conditions. ( A ) Following BMP2 knockdown, significant upregulation of Smad1, Smad4, Smad5, and p38 mRNA expression was observed, with respective increases of 110%, 61%, 36%, and 38% compared to control groups ( p < 0.05 for all comparisons). ( B ) Conversely, BMP2 overexpression resulted in marked downregulation of these targets, showing respective reductions of 25%, 29%, 16%, and 22% in mRNA expression levels ( p < 0.05 for all measurements). All experimental data demonstrated statistically significant differences between treatment groups and corresponding controls. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Experiments in this study were conducted with the following components: fetal bovine serum (Life-ilab, China); Cell Counting Kit-8 (CCK8) kit (ZomanBio, China); Transwell chambers (Corning, USA); Radio Immunoprecipitation Assay Lysis buffer, anti-mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, and anti-rabbit Smad1/5 polyclonal antibody (Boster Bio, China); anti-rabbit p-Smad1/5 (Bioss, China); anti-rabbit BMP2 polyclonal antibody, anti-rabbit p38 polyclonal antibody, and anti-rabbit p-p38 polyclonal antibody (Wanlei Biology, Liaoning); BMP2 small interfering RNA (si-BMP2), control small interfering RNA (si-NC), BMP2 -OE plasmid, control plasmid, and Transfect-Mate transfection reagent (GenePharma, China); and Quantitative Rea-ltime-PCR (qPCR) primers (Thermo Fisher, USA).

Techniques: Expressing, Knockdown, Control, Over Expression

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Bone morphogenetic protein 6 induces downregulation of pentraxin 3 expression in human granulosa lutein cells in women with polycystic ovary syndrome

doi: 10.1007/s10815-023-02972-z

Figure Lengend Snippet: ALK2/ALK3 type I receptors and BMPR2 type II receptor mediate BMP6-induced phosphorylation of SMAD1/5/8 in KGN cells. a KGN cells were pretreated for 1 h with DMSO, DM (10 µM), DMH-1 (0.25 µM), or SB431542 (10 µM), and cells were then treated with 100 ng/mL of BMP6 for another 6 h. The levels of phosphorylated SMAD1/5/8 protein levels were examined using Western blot analysis. b KGN cells were transfected with siCtrl, siALK2, siALK3, or siALK6 for 24 h, and cells were then treated with 100 ng/mL of BMP6 for another 1 h. The phosphorylated SMAD1/5/8 protein levels were examined using Western blot analysis. c KGN cells were transfected with siCtrl, siALK2, siALK3 or combined siALK2 and siALK3 for 24 h, and cells were then treated with 100 ng/mL of BMP6 for another 1 h. The phosphorylated SMAD1/5/8 protein levels were examined using Western blot analysis. d–f KGN cells were transfected with siCtrl, siBMPR2 ( d ), siACVR2A ( e ), or siACVR2B ( f ) for 48 h, and cells were then treated with 100 ng/mL of BMP6 for another 6 h. The phosphorylated SMAD1/5/8 protein levels were examined using Western blot analysis. g–h KGN cells were transfected with siCtrl, combined siBMPR2 and siACVR2A, combined siBMPR2 and siACVR2B, or combined siACVR2A and siACVR2B for 48 h, and cells were then treated with 100 ng/mL BMP6 for another 6 h. The phosphorylated SMAD1/5/8 protein levels were examined using Western blot analysis

Article Snippet: Polyclonal rabbit anti-phospho-SMAD1 (Ser463/465) antibody , Cell Signaling Technology (Beverly, MA).

Techniques: Phospho-proteomics, Western Blot, Transfection

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Bone morphogenetic protein 6 induces downregulation of pentraxin 3 expression in human granulosa lutein cells in women with polycystic ovary syndrome

doi: 10.1007/s10815-023-02972-z

Figure Lengend Snippet: SMAD1 and SMAD5 mediate the suppressive effect of BMP6 on PTX3 expression in KGN cells. a–c KGN cells were transfected with 25 nM siCtrl, 25 nM SMAD1 siRNA (siSMAD1) ( a ), 25 nM SMAD5 siRNA (siSMAD5) ( b ), or 25 nM SMAD8 siRNA (siSMAD8) ( c ) for 24 h. The mRNA levels of SMAD1, SMAD5, and SMAD8 were examined using RT-qPCR. d–f KGN cells were transfected with 25 nM siCtrl, 25 nM siSMAD1 ( d ), 25 nM siSMAD5 ( e ), or 25 nM siSMAD8 ( f ) for 24 h, and cells were then treated with 100 ng/mL BMP6 for another 6 h. The mRNA levels of PTX3 were examined using RT-qPCR. g–i KGN cells were transfected with 25 nM siCtrl, concomitant 25 nM siSMAD1 and siSMAD5 ( g ), concomitant 25 nM siSMAD1 and siSMAD8 (h), or concomitant 25 nM siSMAD5 and siSMAD8 ( i ) for 24 h and then treated with 100 ng/mL BMP6 for 6 h. The mRNA levels of PTX3 were examined using RT-qPCR. The results are expressed as means ± SEMs of at least three independent experiments. Different letters indicate significant differences ( P < 0.05)

Article Snippet: Polyclonal rabbit anti-phospho-SMAD1 (Ser463/465) antibody , Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Bone morphogenetic protein 6 induces downregulation of pentraxin 3 expression in human granulosa lutein cells in women with polycystic ovary syndrome

doi: 10.1007/s10815-023-02972-z

Figure Lengend Snippet: Antibodies and reagents

Article Snippet: Polyclonal rabbit anti-phospho-SMAD1 (Ser463/465) antibody , Cell Signaling Technology (Beverly, MA).

Techniques: Recombinant