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