Journal: PLoS ONE

Article Title: Annexin A1 Induces Skeletal Muscle Cell Migration Acting through Formyl Peptide Receptors

doi: 10.1371/journal.pone.0048246

Figure Lengend Snippet: Cell surface (a) and extracellular (b) ANXA1 from C2C12 cells in GM (0 differentiation day) and after exposure for the indicated times (3, 5, and 7 differentiation days) to DM was analyzed by Western blot with anti-ANXA1 (a, b) antibody. Total cell protein extracts were analyzed by Western blot with anti-ANXA1 (c) and with anti-MyoD (d), anti-Myogenin (e), and anti-MyHC (f) antibodies to assess myogenic differentiation rate. The protein bands were normalized on tubulin levels (g). The data are representative of 5 experiments with similar results.

Article Snippet: Treatments with ionomycin (1 mM) and/or fMLP (50 nM; Sigma-Aldrich), ANXA1 N-terminal peptide Ac2-26 (100 nM; Tocris Biosciences), cyclosporine H (CsH; 500 nM; Alexis-Biochemicals) were carried out by adding the appropriate concentrations of each substance into the cuvette in Ca 2+ -free HBSS/0.5 mM EDTA buffer.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Annexin A1 Induces Skeletal Muscle Cell Migration Acting through Formyl Peptide Receptors

doi: 10.1371/journal.pone.0048246

Figure Lengend Snippet: ( A ) Results for control, fMLP, Ac2-26, CsH, fMLP + CsH and Ac2-26+ CsH are reported as means of three experiments, measuring individual cell migrations at different times. Bars represent standard errors. ( B ) Cytosolic, cell surface and extracellular ANXA1 from C2C12 cells after exposure for the indicated time (16 h) to fMLP, Ac2-26, CsH, Ac2-26+ CsH and fMLP + CsH were analyzed by Western blot with anti-ANXA1 and anti-tubulin antibodies. The data are representative of 5 experiments with similar results. *** <0.001 vs control; §§§ <0.001 vs Ac 2-26 or fMLP.

Article Snippet: Treatments with ionomycin (1 mM) and/or fMLP (50 nM; Sigma-Aldrich), ANXA1 N-terminal peptide Ac2-26 (100 nM; Tocris Biosciences), cyclosporine H (CsH; 500 nM; Alexis-Biochemicals) were carried out by adding the appropriate concentrations of each substance into the cuvette in Ca 2+ -free HBSS/0.5 mM EDTA buffer.

Techniques: Control, Western Blot

Journal: PLoS ONE

Article Title: Annexin A1 Induces Skeletal Muscle Cell Migration Acting through Formyl Peptide Receptors

doi: 10.1371/journal.pone.0048246

Figure Lengend Snippet: Proteins identified as possible Ac2-26 partners by chemical proteomics.

Article Snippet: Treatments with ionomycin (1 mM) and/or fMLP (50 nM; Sigma-Aldrich), ANXA1 N-terminal peptide Ac2-26 (100 nM; Tocris Biosciences), cyclosporine H (CsH; 500 nM; Alexis-Biochemicals) were carried out by adding the appropriate concentrations of each substance into the cuvette in Ca 2+ -free HBSS/0.5 mM EDTA buffer.

Techniques:

Journal: PLoS ONE

Article Title: Annexin A1 Induces Skeletal Muscle Cell Migration Acting through Formyl Peptide Receptors

doi: 10.1371/journal.pone.0048246

Figure Lengend Snippet: Cultured murine C2C12 myoblasts fixed and labeled with fluorescent antibody against ANXA1 and with FITC-conjugated Phalloidin in Ca 2+ -free conditions (a, c, e, g) and in a medium containing 2 mM Ca 2+ (b, d, f, h). (B) An increase of intracellular Ca 2+ levels leads to ANXA1 relocation to the plasma membrane (a); at high Ca 2+ concentrations actin filaments (green) are significantly damaged by high intracellular increase of Ca 2+ level (e): in this condition ANXA1(red) is strikingly shifted to the plasma membrane (d). Bar = 25 µm.

Article Snippet: Treatments with ionomycin (1 mM) and/or fMLP (50 nM; Sigma-Aldrich), ANXA1 N-terminal peptide Ac2-26 (100 nM; Tocris Biosciences), cyclosporine H (CsH; 500 nM; Alexis-Biochemicals) were carried out by adding the appropriate concentrations of each substance into the cuvette in Ca 2+ -free HBSS/0.5 mM EDTA buffer.

Techniques: Cell Culture, Labeling, Clinical Proteomics, Membrane