Journal: Scientific Reports

Article Title: Human peripheral blood mononuclear cells display a temporal evolving inflammatory profile after myocardial infarction and modify myocardial fibroblasts phenotype

doi: 10.1038/s41598-023-44036-3

Figure Lengend Snippet: Antibodies and dies for flow cytometry.

Article Snippet: Cells were then stained with 50 µL of anti-P2Y11 AF405 secondary antibody (goat anti-rabbit, 1:250, ThermoFisher Scientific, Waltham, MA, USA) and 2 µL of anti-alpha-smooth muscle actin-PE (clone 1A4, Bio-techne, Mineapolis, MI, USA) during 1 h at 4 °C.

Techniques: Cytometry, Concentration Assay

Journal: Respiratory research

Article Title: Intraluminal chloride regulates lung branching morphogenesis: involvement of PIEZO1/PIEZO2.

doi: 10.1186/s12931-023-02328-2

Figure Lengend Snippet: Fig. 6 Molecular effect of intraluminal chloride and GsMTx4 in smooth muscle cells. a-c Upper panel represents the main cumulative effect of intraluminal chloride concentration ([Cl−]) and medium supplementation with and without GsMTx4. a Examples of representative blots are shown. b, c Protein expression levels for b myosin light chain 2 (MLC2), and c alpha-smooth muscle actin (α-SMA) are quantified. d–f Lower panel shows the additional effect of PIEZO1/2 inhibition after intraluminal injection of Cl− channels inhibitors: cystic fibrosis transmembrane conductance regulator inhibitor172 (CFTRinh) to CFTR and; calcium-dependent Cl− channel inhibitor A01 (CaCCinh) to CaCCs. d Examples of representative blots are shown. e–f Relative expression levels of e MLC2, and f α-SMA are displayed. 143 mM Cl− and standard solution (SS) represent the control condition for [Cl−] and Cl− channels inhibitors, respectively. White and dotted rectangles represent the medium supplementation with and without GsMTx4, respectively. Each lane represents a pooled-tissue sample, and the relative expression levels were determined against GAPDH. n ≥ 4 were used per antibody/condition. Results are presented as mean ± SD. Symbols indicate the main effects and non-redundant interactions of the two-way ANOVA. p < α0.0001, β0.001, γ0.01, µ0.05

Article Snippet: Blots were blocked in 5% bovine serum albumin and probed with primary antibodies to ghrelin (1:250, ON, 4 oC; Cat No. sc10368, Santa Cruz Biotechnology Inc., USA), bombesin (1:250, ON, 4 oC; Cat No. H00002922-MO3, Novus Biologicals, USA), PIEZO1 (1:250, ON, 4 oC; Cat No. HPA047185, Sigma-Aldrich, USA), PIEZO2 (1:250, ON, 4 oC; Cat No. HPA040616, Sigma-Aldrich, USA), myosin light chain 2 (MLC2, 1∶250; ON, 4 oC; Cat No. #3672, Cell Signaling Technology Inc., USA), and alpha-smooth muscle actin (α-SMA, 1:500, ON, 4 oC; Cat No. NBP2-33006, Novus Biologicals, USA) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Expressing, Inhibition, Injection, Control

Journal: Respiratory research

Article Title: Intraluminal chloride regulates lung branching morphogenesis: involvement of PIEZO1/PIEZO2.

doi: 10.1186/s12931-023-02328-2

Figure Lengend Snippet: Fig. 7 Schematic representation of the role of intraluminal chloride concentrations in fetal branching morphogenesis. Increasing concentrations of intraluminal [Cl−] leads to increased branching morphogenesis and expression of neuroendocrine products, PIEZO channels, and smooth muscle markers. Pharmacological inhibition experiments show that these effects require PIEZO and Cl− channel activation. α-SMA, alpha-smooth muscle actin; CaCCinh calcium-dependent Cl− channel inhibitorA01; CFTRinh, cystic fibrosis transmembrane conductance regulator inhibitor172; MLC2, myosin light chain 2; SS, standard solution

Article Snippet: Blots were blocked in 5% bovine serum albumin and probed with primary antibodies to ghrelin (1:250, ON, 4 oC; Cat No. sc10368, Santa Cruz Biotechnology Inc., USA), bombesin (1:250, ON, 4 oC; Cat No. H00002922-MO3, Novus Biologicals, USA), PIEZO1 (1:250, ON, 4 oC; Cat No. HPA047185, Sigma-Aldrich, USA), PIEZO2 (1:250, ON, 4 oC; Cat No. HPA040616, Sigma-Aldrich, USA), myosin light chain 2 (MLC2, 1∶250; ON, 4 oC; Cat No. #3672, Cell Signaling Technology Inc., USA), and alpha-smooth muscle actin (α-SMA, 1:500, ON, 4 oC; Cat No. NBP2-33006, Novus Biologicals, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Inhibition, Activation Assay

Journal: Neurotrauma Reports

Article Title: Early Transcutaneous Tibial Nerve Stimulation Acutely Improves Lower Urinary Tract Function in Spinal Cord Injured Rats

doi: 10.1089/neur.2021.0058

Figure Lengend Snippet: Assessment of bladder hypertrophy four weeks after spinal cord injury. ( A ) Bladder weight. ( B ) Quantification of the muscle to collagen ratio obtained by Masson Trichrome staining. ( C ) Representative Masson Trichrome staining of bladder tissue. Muscle is stained red, collagen blue. ( D–F ) Quantification of three muscle proteins by Western blot (WES): myosin heavy chain 11 (MYH11; 182 kDa), alpha-smooth muscle actin (SMA; 48 kDa), and calponin (45 kDa) in sham and stimulated groups. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. * p < 0.05.

Article Snippet: The primary antibodies used for detection were mouse primary antibodies against alpha smooth muscle actin (αSMA) (Novus Biologicals, Centennial, CO, NBP2-33006, 1:100), calponin (Sigma-Aldrich, C2687, 1:400), and myosin heavy chain 11 (MYH11) (Santa Cruz, sc-6956, 1:10).

Techniques: Staining, Western Blot

Journal: Cells

Article Title: Neonatal Porcine Germ Cells Dedifferentiate and Display Osteogenic and Pluripotency Properties

doi: 10.3390/cells10112816

Figure Lengend Snippet: Antibodies used for immunofluorescence and immunocytochemistry.

Article Snippet: Mouse anti-ASM , Novus Biological , NBP1-33006 , 1:200.

Techniques: Immunofluorescence, Immunocytochemistry