function-blocking antibody to integrin αvβ3  (Merck KGaA)

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: Reverse transcription PCR showing the expression of ( A ) β3 integrin and ( B ) β1 integrin in myoblasts and myosphere cells. Representative FACS profiles showing β3 integrin expression by ( C ) myosphere cells and ( D ) myoblasts. Empty peaks represent the unstained controls, filled peaks cells incubated with PE conjugated β3 integrin antibody. ( E ) Graph showing the number of cells adhering/field to αVβ3 integrin versus cells tested, myoblasts are shown in white and myosphere cells in black. These results were pooled from two independent experiments. Error bars display ± s.e.m.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Reverse Transcription, Expressing, Incubation

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: Representative FACS profiles showing Sca-1 expression by ( A ) myosphere cells and ( B ) myoblasts. Empty peaks represent unstained controls, filled peaks cells incubated with FITC conjugated Sca-1 antibody. ( C ) Graph showing percent of cells adhering/field to αVβ3 integrin versus cells incubated with antibody, represented are myoblasts (white), myosphere cells incubated with: no antibody (black), Sca-1 E13 antibody (light gray), Sca-1 D7 antibody (medium gray), and β1 integrin antibody (dark gray). Each of the Sca-1 antibody results was pooled from two independent experiments; the β1 antibody results were pooled from three independent experiments. Error bars display ± s.e.m. *Indicates P = 0.029, ***Indicates P <0.0001 both by unpaired Student's t test.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Expressing, Incubation

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: ( A ) Representative FACS profile showing Sca-1 expression in NIH3T3 cells. Empty peak represents the unstained control; filled peak represents cells incubated with FITC conjugated Sca-1 antibody. ( B ) Graph showing the percent of cells adhering/field to αVβ3 integrin versus cells tested, represented are NIH3T3 cells with no antibody (white), Sca-1 antibody (light gray), transduced with control shRNA vector (dark gray), and transduced with shRNA Sca-1 knockdown vector (black). FACS profiles show Sca-1 expression in NIH3T3 cells transduced by the ( C ) shRNA Sca-1 knockdown vector and ( D ) control shRNA vector. Empty peaks represent the unstained control; filled peaks represent cells incubated with APC conjugated Sca-1 antibody. The results in this figure are pooled from 3 independent experiments. Error bars display ± s.e.m. *** P <0.0001 by unpaired Student's t test.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Expressing, Control, Incubation, Transduction, shRNA, Plasmid Preparation, Knockdown

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: ( A ) Bicistronic lentiviral vector used to transduce primary myoblasts. ( B ) FACS profile showing Sca-1 expression by transduced myoblasts. Empty peak represents the unstained control, filled peak myoblasts incubated with FITC conjugated Sca-1 antibody. ( C ) Western showing the expression of Sca-1 in untransduced, control transduced, and Sca-1 transduced myoblasts. ( D ) Graph showing the percent of cells adhering/field to αVβ3 integrin versus cells tested, represented are myoblasts (white), myoblasts transduced by the control vector (light gray), myoblasts transduced by the Sca-1 vector (medium gray, MOI = 5) and (dark gray, MOI = 50), and myosphere cells (black). Results were pooled from 6 independent adhesion assays (3 for cell transduced at an MOI = 5, and 3 for those transduced at a MOI = 50). Error bars display ± s.e.m. *** P <0.0001 by unpaired Student's t test.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Plasmid Preparation, Transduction, Expressing, Control, Incubation, Western Blot

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: Representative FACS profiles showing the expression of ( A ) αV integrin and ( B ) β3 integrin in NIH3T3 cells. Empty peaks represent the unstained control; filled peaks represent cells incubated with PE conjugated αV and β3 integrin antibodies. ( C ) FACS profile of NIH3T3 cells after transduction with the β3 integrin vector. Empty peak represents the unstained control; filled peak represents cells incubated with APC conjugated β3 integrin antibody. ( D ) Graph showing the percent of cells adhering/field to αVβ3 integrin versus cells tested, represented are NIH3T3 cells (black), NIH3T3 cells expressing β3 integrin (dark gray), NIH3T3 cells expressing neither Sca-1 or β3 integrin (white). ( E ) Graph showing the percent of cells adhering/field to αVβ3 integrin versus cells tested, represented are NIH3T3 cells expressing both Sca-1 and β3 integrin incubated with: no antibodies (black), Sca-1 antibody (dark gray), β3 integrin antibody (light gray), both Sca-1 and β3 integrin antibodies (white). The results in D,E were pooled from 3 independent experiments. Error bars display ± s.e.m. *** P <0.0001 by unpaired Student's t test.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Expressing, Control, Incubation, Transduction, Plasmid Preparation

Journal: Biology Open

Article Title: Sca-1 is involved in the adhesion of myosphere cells to αVβ3 integrin

doi: 10.1242/bio.20121222

Figure Lengend Snippet: ( A ) Myospheres generated from β3 integrin null mice. FACS profiles showing the expression of ( B ) β3 integrin and ( C ) Sca-1 by β3 null myospheres. Empty peaks represent the unstained controls; filled peaks represents cells incubated with PE conjugated to β3 integrin antibody and APC conjugated to Sca-1 antibody, respectively. ( D ) Graph showing the percent of cells adhering/field to αVβ3 integrin versus cells tested; represented are myosphere cells isolated from wild type (black), Sca-1 null (dark gray), and β3 integrin null (light gray) mice. These results were pooled from 2 independent experiments. Error bars display ± s.e.m. *** P <0.0001 by unpaired Student's t test. Measure bar shown in A represents 100 µm.

Article Snippet: As an additional control some of the αVβ3 integrin coated wells were incubated with 10 μg/ml of an antibody that functionally blocks αVβ3 integrin (clone LM609, Millipore; Billerica, MA, USA).

Techniques: Generated, Expressing, Incubation, Isolation