Journal: Biology

Article Title: BMP-2-Driven Osteogenesis: A Comparative Analysis of Porcine BMSCs and ASCs and the Role of TGF-β and FGF Signaling.

doi: 10.3390/biology14060610

Figure Lengend Snippet: Figure 6. Receptor expressions of ALK2, ALK4, ALK6, ALK5, ALK7, BMPR-II in pASC. Grouped representation of the respective receptor expression in the course of osteogenic differentiation (OM +/−BMP-2). From day 19, there is a significant induction of ALK 2, ALK 6, and ALK 5 with the addition of BMP-2. BMPR-II expression in the OM group decreased in OM and tended to stay increased under BMP-2 supplementation from day 19, but was not considered significant (* p ≤0.05, ** p ≤0.01; n = 6, BMP-2 450 ng/mL).

Article Snippet: The respective conjugated antibodies were used for the expressions of ALK3 (Cat. No.: AF436), ALK 5 (Cat. No.: FAB5871), ALK6 (Cat. No.: FAB5051A), TGF-β2-RII (Cat. No.: FAB532P), ALK7 (Cat. No.: FAB77491A), ALK2 (Cat. No.: AF637), ALK4 (Cat. No.: MAB2221), and BMPR-II (Cat. No.: AF811) (by R&D Systems, Minneapolis, MN, USA), and the pASCs and pBMSCs were compared for their expressions of the specific surface antigens CD45 (Cat. No.: MCA1568GA, BioRad, Hercules, CA, USA), HLA-DR (human leukocyte antigen–antigen D-related surface molecule) (Cat. No.: MCA2314F, Bio-Rad, Hercules, CA, USA), CD29 (Cat. No.: 561,496, BD Pharmingen, Franklin Lakes, NJ, USA), CD79alpha (Bio-Rad, Cat. No.: MCA2538GA), CD14 (Cat. No.: MCA1568GA, Bio-Rad, Hercules, CA, USA), CD31 (Cat. No.: AF3387, R&D Systems, Minneapolis, MN, USA), CD105 (Cat. No.: NB110-58718APC, Novus Biologicals, Minneapolis, MN, USA), CD26 (, Cat. No.: NB600-552APC, Novus Biologicals, Minneapolis, MN, USA), CD73 (, Cat. No.: AF4488, R&D Systems, Minneapolis, MN, USA), CD90 (Cat. No.: 559,869, BD Pharmingen, Franklin Lakes, NJ, USA), CD34 (Cat. No.: 81289, abcam, Cambridge, UK), and CD44 (Cat. No.: 5531, BD Pharmingen, Franklin Lakes, NJ, USA).

Techniques: Expressing

Journal: PLoS ONE

Article Title: BMP-7 induces apoptosis in human germinal center B cells and is influenced by TGF-β receptor type I ALK5

doi: 10.1371/journal.pone.0177188

Figure Lengend Snippet: (A-C) GC B cells were obtained by immunomagnetic bead separation and co-cultured with HK cells in the presence of CD40L/IL-21 with or without BMP-7 and apoptosis was measured by TUNEL assay. (A) Cells were cultured for up to 3 days and analyzed by flow cytometry. Shown is one representative of 2 donors. (B) After 2 days in culture, TUNEL staining (green) and Hoechst staining (blue) was detected by confocal microscopy. Representative images from one of three independent experiments are presented. Scale bar represents 30 μm. (C) The ALK 2/3 and ALK 4/5/7 selective inhibitors, LDN193189 and SB431542, respectively, were added to the cultures as specified and apoptosis was measured by flow cytometry at day 2. Mean ±SEM, n = 4, (D) Single cell suspensions from human tonsils were stained with lineage markers and anti-ALK4, anti-ALK5 or anti-ALK7 antibodies, and analyzed by flow cytometry. Shown are histogram overlays of receptor expression as compared to an irrelevant control. All experiments were repeated at least twice. Statistical testing was performed against CD40L/IL-21 condition. * p < 0.05; two-tailed, paired Student’s t -test.

Article Snippet: The following primary antibodies were used: biotinylated anti-ActRIIa (BAF340), -ActRIIb (BAF339), -BMPRII (BAF811),—ALK2 (BAF637), -ALK3 (BAF820), -ALK4 (BAF222) -ALK5 (BAF 3025), -ALK6 (BAF505), biotinylated goat IgG (BAF108) (R&D Systems, MN, USA).

Techniques: Cell Culture, TUNEL Assay, Flow Cytometry, Staining, Confocal Microscopy, Expressing, Control, Two Tailed Test

Journal: PLoS ONE

Article Title: BMP-7 induces apoptosis in human germinal center B cells and is influenced by TGF-β receptor type I ALK5

doi: 10.1371/journal.pone.0177188

Figure Lengend Snippet: Mino cells were transduced with truncated ALK5 or truncated ALK4. (A) The cells were stained with biotinylated anti-ALK5, followed by streptavidin PE or by biotinylated anti-ALK4, followed by streptavidin APC, and analyzed by flow cytometry. Receptor expression is compared in GFP + or mCherry + transduced cells vs. GFP - /mCherry - non-transduced cells. (B-C) The transduced Mino cells were cultured in serum free media (X-VIVO 15) over night and then left in medium alone (unstim) or stimulated with BMP-2, BMP-7 or TGF-β for 60 min, before detection of phosphorylated (p-) Smad 1/5 or p-Smad 2/3 by flow cytometry. (B) One representative experiment showing p-SMAD1/5 vs. GFP in truncated ALK5-2A-GFP expressing cells. (C) BMP- or TGF-β-induced phosphorylation is shown relative to unstimulated cells, using arcsinh transformation of median fluorescence intensity data. Mean ± SEM, n = 5. (D-E): Transduced Mino cells were cultured in X-VIVO 15 and left unstimulated or stimulated with TGF-β or BMP-7 for 72 hours and stained for active caspase-3 before analysis by flow cytometry. Shown here is active caspase-3 staining of control cells and transduced cells for (D) one representative experiment and (E) mean ± SEM, n = 3. * p < 0.05; two-tailed, paired Student’s t -test.

Article Snippet: The following primary antibodies were used: biotinylated anti-ActRIIa (BAF340), -ActRIIb (BAF339), -BMPRII (BAF811),—ALK2 (BAF637), -ALK3 (BAF820), -ALK4 (BAF222) -ALK5 (BAF 3025), -ALK6 (BAF505), biotinylated goat IgG (BAF108) (R&D Systems, MN, USA).

Techniques: Transduction, Staining, Flow Cytometry, Expressing, Cell Culture, Phospho-proteomics, Transformation Assay, Fluorescence, Control, Two Tailed Test

Journal: Anticancer research

Article Title: Potential roles of activin in head and neck squamous cell carcinoma progression in epithelial-mesenchymal transition, metastasis, and mortality

doi: 10.21873/anticanres.16733

Figure Lengend Snippet: The canonical activin A pathway. Activin A is composed of inhibin βA subunits (βA) and binds to activin receptor type II and IIB (ACVR2/2B). Inhibin, composed of a βA and α subunit, competitively binds and sequesters ACVR2/2B, ultimately inhibiting the activin axis. Contrariwise, activin binding ultimately forms a Smad transcriptions complex (comprised of Smad 2, 3 and 4), the phosphorylation of activin receptor type IB (ACVR1B) subsequently stimulating and activating the Smad transcription complex, thereby eliciting downstream cellular behaviors such as proliferation inhibition, apoptosis, and epithelial mesenchymal transition. Of note, activin may have proliferative effects outside of this axis, as described in the introduction and discussion.

Article Snippet: ACVR1B (MAB222) , Monoclonal , R & D Systems , 1:200.

Techniques: Binding Assay, Phospho-proteomics, Inhibition

Journal: Anticancer research

Article Title: Potential roles of activin in head and neck squamous cell carcinoma progression in epithelial-mesenchymal transition, metastasis, and mortality

doi: 10.21873/anticanres.16733

Figure Lengend Snippet: Immunohistochemistry expression of inhibin subunits (INHA, INHBA, INHBB) and activin receptors (ACVR1B, ACVR2, ACVR2B) in five normal, 15 oral premalignant (OPL) and 12 HNSCC tumor tissue samples. Chi-square tests were employed for analysis with p <0.05 being significant; diffuse and focal positivity were scored as positive. Premalignant and malignant lesions demonstrated a statistically significant increase in the prevalence of ligand inhibin βA (INHBA) (χ 2 (2, N = 32) = 18.98, p < .0001) (Row 6) as well as ACVR1B (χ 2 (2, N = 32) = 11.52, p < .0032) (Row 11). There was also a decreased prevalence of ACVR2B among pre-malignant and malignant lesions in comparison to normal mucosa (χ 2 (2, N = 32) = 0.0018, p < .0018) (Row 13).

Article Snippet: ACVR1B (MAB222) , Monoclonal , R & D Systems , 1:200.

Techniques: Immunohistochemistry, Expressing, Comparison

Journal: Anticancer research

Article Title: Potential roles of activin in head and neck squamous cell carcinoma progression in epithelial-mesenchymal transition, metastasis, and mortality

doi: 10.21873/anticanres.16733

Figure Lengend Snippet: Immunohistochemistry

Article Snippet: ACVR1B (MAB222) , Monoclonal , R & D Systems , 1:200.

Techniques: