smad2 3  (Cell Signaling Technology Inc)

Journal: Cells

Article Title: Cytokine Receptor-like Factor 1 (CRLF1) and Its Role in Osteochondral Repair

doi: 10.3390/cells13090757

Figure Lengend Snippet: Chondrogenic differentiation of human BMSCs lacking CRLF1 or overexpressing CRLF1. ( A ) Alcian blue staining of sections of BMSC pellets transfected with control siRNA (siCon) or siRNA specific for CRLF1 (siCRLF1) after 6 days in chondrogenic differentiation medium; sections of BMSC pellet transduced with empty AAV (AAV) or transduced with AAV containing CRLF1 (AAV-CRLF1) after 6 days in chondrogenic differentiation medium. Bar, 100 µm. ( B ) Phosphorylated (p-Smad) and total Smad2 and Smad3 immunoblots of lysates from BMSCs transduced with empty AAV (AAV) or from BMSCs transduced with AAV-CRLF1. Cells were treated for 60 min with transforming growth factor beta 1 (TGFβ1).

Article Snippet: After blocking with a solution of low-fat milk protein, blotted proteins were immunostained with primary antibodies specific for β-actin (#3700, Cell Signaling Technology, Danvers, MA, USA), CRLF1 (ab211438, Abcam, Boston, MA, USA) or CLC (ab154798, Abcam), total STAT3 (#9139, Cell Signaling Technology), phosphorylated STAT3 (#9145, Cell Signaling Technology), total Smad2 and 3, phosphorylated Smad 2, and phosphorylated Smad3 (SMAD2/3 Antibody Sampler Kit #12747, Cell Signaling Technology), and then peroxidase-conjugated secondary antibody (Thermo Scientific Pierce, Waltham, MA, USA).

Techniques: Staining, Transfection, Control, Transduction, Western Blot

Journal: Burns & Trauma

Article Title: miR-125b-5p delivered by adipose-derived stem cell exosomes alleviates hypertrophic scarring by suppressing Smad2

doi: 10.1093/burnst/tkad064

Figure Lengend Snippet: Smad2 plays an important role in hypertrophic scarring. ( a ) H&E staining results of tissue from normal skin and hypertrophic scars. Scale bars: 1 mm or 500 μm. ( b ) Masson staining results of tissue from normal skin and hypertrophic scars. Scale bars: 1 mm or 500 μm. ( c ) α-SMA immunohistochemical staining results of tissue from normal skin and hypertrophic scars. Scale bar: 1 mm or 500 μm. ( d ) Smad2 immunohistochemical staining results of tissue from normal skin and hypertrophic scars. Scale bars: 1 mm or 500 μm. H&E hematoxylin and eosin, α-SMA α-smooth muscle actin

Article Snippet: The membrane was then blocked with 5% skim milk for 1 h at room temperature in TBST and probed with the following antibodies: COL1 (1 : 1000, Abcam, Cambridge, UK), COL3 (1 : 1000, Abcam, Cambridge, UK), α-SMA (1 : 1000, CST, USA), CD9, CD63 (1 : 1000, Proteintech, China), Smad2 antibody sampling kit (1 : 1000, CST) and β-actin (1 : 1000, Xi’an Johnson & Johnson) and incubated overnight at 4°C.

Techniques: Staining, Immunohistochemical staining

Journal: Burns & Trauma

Article Title: miR-125b-5p delivered by adipose-derived stem cell exosomes alleviates hypertrophic scarring by suppressing Smad2

doi: 10.1093/burnst/tkad064

Figure Lengend Snippet: ADSC exosomes downregulate the expression of Smad2. ( a ) PCR results of Smad2 expression in HSFs treated with ADSC Exo and PBS, * * p < 0.01. ( b ) Western blot results of Smad2 and p-Smad2 expression in HSFs treated with ADSC Exo and PBS, * * p < 0.01. ( c ) Smad2 immunofluorescence staining results of HSFs treated with ADSC Exo and PBS, blue: DAPI, red: PKH26, green: Smad2, * * p < 0.01. Scale bar: 125 μm. ADSC adipose-derived stem cells, Exo exosomes, PBS phosphate-buffered saline, HSFs hypertrophic scar fibroblasts

Article Snippet: The membrane was then blocked with 5% skim milk for 1 h at room temperature in TBST and probed with the following antibodies: COL1 (1 : 1000, Abcam, Cambridge, UK), COL3 (1 : 1000, Abcam, Cambridge, UK), α-SMA (1 : 1000, CST, USA), CD9, CD63 (1 : 1000, Proteintech, China), Smad2 antibody sampling kit (1 : 1000, CST) and β-actin (1 : 1000, Xi’an Johnson & Johnson) and incubated overnight at 4°C.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Derivative Assay, Saline

Journal: Burns & Trauma

Article Title: miR-125b-5p delivered by adipose-derived stem cell exosomes alleviates hypertrophic scarring by suppressing Smad2

doi: 10.1093/burnst/tkad064

Figure Lengend Snippet: ADSC Exo downregulate the expression of Smad2 by delivering miR-125b-5p. ( a ) Hotmap showing miRNA microarray results of ADSC Exo. ( b ) PCR results of the expression of 5 miRNAs (miR-125b-5p, miR-10a-5p, miR-23a-3p, miR-21-5p and miR-92a-3p) in ADSC Exo. ( c ) PCR results indicating miR-125b-5p expression in PBS- and ADSC Exo-treated HSFs, * * p < 0.01. ( d ) Structure and luciferase results of the dual luciferase reporter gene, * * p <0.01. ( e ) PCR results indicating miR-125b-5p expression in negative control (NC), miR-125b-5p expression mimic- and inhibitor-treated HSFs, * * p < 0.01. ( f ) Western blot results of α-SMA, COL1, COL3, p-Smad2 and Smad2 expression in NC, miR-125b-5p expression mimic- and inhibitor-treated HSFs. ADSC adipose-derived stem cells, Exo exosomes, PBS phosphate-buffered saline, H&E hematoxylin and eosin, HSFs hypertrophic scar fibroblasts

Article Snippet: The membrane was then blocked with 5% skim milk for 1 h at room temperature in TBST and probed with the following antibodies: COL1 (1 : 1000, Abcam, Cambridge, UK), COL3 (1 : 1000, Abcam, Cambridge, UK), α-SMA (1 : 1000, CST, USA), CD9, CD63 (1 : 1000, Proteintech, China), Smad2 antibody sampling kit (1 : 1000, CST) and β-actin (1 : 1000, Xi’an Johnson & Johnson) and incubated overnight at 4°C.

Techniques: Expressing, Microarray, Luciferase, Negative Control, Western Blot, Derivative Assay, Saline

Journal: Burns & Trauma

Article Title: miR-125b-5p delivered by adipose-derived stem cell exosomes alleviates hypertrophic scarring by suppressing Smad2

doi: 10.1093/burnst/tkad064

Figure Lengend Snippet: ADSC Exos alleviate fibrosis and decrease the expression of Smad2 in a mouse fibrosis model. ( a ) H&E staining results of ADSC Exo or PBS-treated bleomycin-induced fibrosis. Scale bars: 1 mm or 400 μm. ( b ) Masson staining results of ADSC Exo or PBS-treated bleomycin-induced fibrosis. Scale bars: 1 mm or 400 μm. ( c ) Smad2 immunofluorescence and immunohistochemical staining results of ADSC Exo or PBS-treated bleomycin-induced fibrosis, blue: DAPI, green: Smad2. Scale bars: 500 or 400 μm. ADSC adipose-derived stem cells, Exo exosomes, PBS phosphate-buffered saline, H&E hematoxylin and eosin

Article Snippet: The membrane was then blocked with 5% skim milk for 1 h at room temperature in TBST and probed with the following antibodies: COL1 (1 : 1000, Abcam, Cambridge, UK), COL3 (1 : 1000, Abcam, Cambridge, UK), α-SMA (1 : 1000, CST, USA), CD9, CD63 (1 : 1000, Proteintech, China), Smad2 antibody sampling kit (1 : 1000, CST) and β-actin (1 : 1000, Xi’an Johnson & Johnson) and incubated overnight at 4°C.

Techniques: Expressing, Staining, Immunofluorescence, Immunohistochemical staining, Derivative Assay, Saline