41d10 cell signaling technology Search Results


41d10 cell signaling technology  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc 41d10 cell signaling technology
41d10 Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Smad1%2F5+(Ser463%2F465)+Rabbit+mAb/pm40057955-290-45-46
Average 96 stars, based on 1 article reviews
41d10 cell signaling technology - by Bioz Stars, 2026-09
96/100 stars
  Buy from Supplier
anti phospho smad1 5 41d10 rabbit antibodies  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc anti phospho smad1 5 41d10 rabbit antibodies
Anti Phospho Smad1 5 41d10 Rabbit Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Smad1%2F5+(Ser463%2F465)+Rabbit+mAb/pmc04423117-412-4-8
Average 95 stars, based on 1 article reviews
anti phospho smad1 5 41d10 rabbit antibodies - by Bioz Stars, 2026-09
95/100 stars
  Buy from Supplier
phospho smad1 5 ser463 465  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc phospho smad1 5 ser463 465
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Phospho Smad1 5 Ser463 465, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Smad1+(Ser463%2F465)%2F+Smad5+(Ser463%2F465)%2F+Smad9+(Ser465%2F467)+Rabbit+mAb/pmc11898274-192-10-25
Average 95 stars, based on 1 article reviews
phospho smad1 5 ser463 465 - by Bioz Stars, 2026-09
95/100 stars
  Buy from Supplier
rabbit anti phospho smad1 5 8  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc rabbit anti phospho smad1 5 8
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Rabbit Anti Phospho Smad1 5 8, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Smad1+Antibody/pmc04974506-160-14-29
Average 96 stars, based on 1 article reviews
rabbit anti phospho smad1 5 8 - by Bioz Stars, 2026-09
96/100 stars
  Buy from Supplier
anti phosphosmad1 5 antibody  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc anti phosphosmad1 5 antibody
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Anti Phosphosmad1 5 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Rpb1+CTD+(Ser2%2F5)+Antibody/pmc03493910-172-11-14
Average 93 stars, based on 1 article reviews
anti phosphosmad1 5 antibody - by Bioz Stars, 2026-09
93/100 stars
  Buy from Supplier
anti psmad1 5 41d10 monoclonal antibodies  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti psmad1 5 41d10 monoclonal antibodies
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Anti Psmad1 5 41d10 Monoclonal Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Smad1+(Ser463%2F465)%2F+Smad5+(Ser463%2F465)%2F+Smad9+(Ser465%2F467)+Rabbit+mAb/pmc09708949-94-7-11
Average 96 stars, based on 1 article reviews
anti psmad1 5 41d10 monoclonal antibodies - by Bioz Stars, 2026-09
96/100 stars
  Buy from Supplier
nbp1 06713 rrid ab 1625979 anti phospho smad1 5 41d10  (Novus Biologicals)
94
Novus Biologicals nbp1 06713 rrid ab 1625979 anti phospho smad1 5 41d10
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Nbp1 06713 Rrid Ab 1625979 Anti Phospho Smad1 5 41d10, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/OLLAS+Epitope+Tag+Antibody+(L2)+-+BSA+Free/pm37054707-251-36-33
Average 94 stars, based on 1 article reviews
nbp1 06713 rrid ab 1625979 anti phospho smad1 5 41d10 - by Bioz Stars, 2026-09
94/100 stars
  Buy from Supplier
rabbit anti p mad  (Cell Signaling Technology Inc)
91
Cell Signaling Technology Inc rabbit anti p mad
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
Rabbit Anti P Mad, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Mad-1+Antibody/pm36473687-275-16-20
Average 91 stars, based on 1 article reviews
rabbit anti p mad - by Bioz Stars, 2026-09
91/100 stars
  Buy from Supplier
41d10 rabbit mab antibody cell signaling technology  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc 41d10 rabbit mab antibody cell signaling technology
The interaction IL34-BMP2 modulates <t>SMAD1/5</t> as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.
41d10 Rabbit Mab Antibody Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/41d10+cell+signaling+technology/Phospho-Akt+(Ser473)+Antibody/10__2139_slash_ssrn__3501841-168-65-69
Average 99 stars, based on 1 article reviews
41d10 rabbit mab antibody cell signaling technology - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier
recombinant cynomolgus commd10 protein, his-tagged  (Creative BioMart)
N/A
Recombinant Cynomolgus COMMD10 full length or partial length protein was expressed.http://www.creativebiomart.net/Recombinant-Cynomolgus-COMMD10-Protein-His-tagged-455918.htm
   Buy from Supplier
recombinant mouse cyp2d10 protein  (Creative BioMart)
N/A
Recombinant Mouse CYP2D10 full length or partial length protein was expressed.http://www.creativebiomart.net/Recombinant-Mouse-CYP2D10-Protein-440604.htm
   Buy from Supplier

Image Search Results


The interaction IL34-BMP2 modulates SMAD1/5 as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.

Journal: Theranostics

Article Title: Interleukin-34 orchestrates bone formation through its binding to bone morphogenic proteins

doi: 10.7150/thno.107340

Figure Lengend Snippet: The interaction IL34-BMP2 modulates SMAD1/5 as well as MCSF receptor (MCSFR) phosphorylation and related signaling. (A) Western blot analysis of SMDA1/5 and SMAD2 phosphorylations of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), TGFß (10 ng/mL), MCSF (20 ng/mL), IL34 (20 ng/mL) alone or in corresponding combination. A representative experiment is shown. CT: basic culture medium. (B) Western blot analysis of SMDA1/5 phosphorylation of human MNNG-HOS osteosarcoma cells in the presence of BMP2 (10 ng/mL), IL34 (20 ng/mL) alone or in combination (BMP2+IL34, -) plus the human IL34 blocking antibody (BT34) (100 µg/mL) or its irrelevant isotypic control antibody (ISO) (100 µg/mL). CT: basic culture medium. (C) Western blot analysis of SMDA1/5, similar conditions used in B in the presence of the human IL34 blocking antibody (BT34) (100 µg/mL) or the natural inhibitor of BMP2 called NOGGIN (NOG) (200 ng/mL). (D) Kinetic analysis by Western blot of the potentiating effect of IL34 on BMP2-induced SMAD1/5 phosphorylation at 15 min, 30 min and 60 min with similar corresponding molecules concentrations described in B. (E) Western blot analysis of SMDA1/5 as described in B in the presence of a single concentration of BMP2 (10 ng/mL) in combination with gradual quantities of IL34 (10, 20, 40, 80 and 100 ng/mL). (F) The Alpha SureFire technology (Revvity) was used to quantitatively validate the potentiation effect of IL34 on BMP2 activation of SMAD1/5 phosphorylation. Co-additions of 25 or 50 ng/mL of IL34 increased significantly the phosphorylation of SMAD1/5 induced by the addition of BMP2 at 10 ng/mL. **p<0.01, ***p<0.001. (G) Western blot analysis of SMDA1/5 as described in B with a single concentration of IL34 (20 ng/mL) in combination with gradual quantities of IL34 (5, 10, 20, 40 and 80 ng/mL). (H) Western blot analysis of MCSFR phosphorylation expressed in HEK293 transfected cells in the presence or absence of BMP2 (10 ng/mL), IL34 (20 ng/mL) or in combination (BMP2+IL34, -) plus NOGGIN (NOG) (200 ng/mL). Quantifications of all the Western blots presented in this figure are shown in . All experiments have done at least three times independently.

Article Snippet: Antibodies directed against human Smad1 (D59D7), human Smad2 (D43B4), anti Phospho-Smad1/5 (ser463/465) (41D10), anti-phospho-Smad2 (Ser465/467) (138D), β-Actin (8H10D10) and HRP-conjugated secondary antibodies were purchased from Cell Signalling (Ozyme, Saint Quentin Yvelines, France).

Techniques: Western Blot, Blocking Assay, Control, Concentration Assay, Activation Assay, Transfection