Journal: Biology of Reproduction

Article Title: Effects of vaginal conjugated equine estrogens and ospemifene on the rat vaginal wall and lower urinary tract †

doi: 10.1095/biolreprod.116.144428

Figure Lengend Snippet: Antibody Table.

Article Snippet: Thereafter, membranes were incubated with blocking buffer (tris-buffered saline-Tween 20 with 5% nonfat milk) for 1 h. An affinity-purified rabbit antibody against rat and mouse collagen type 1 (COL1a1) (Merck-Millpore, Darmstadt, Germany) and an ascites-grade mouse antibody against human and rat collagen type 3 (COL3a1) (USBiological, Salem, MA) was used for immunoblotting (Table ).

Techniques:

Journal: Chinese Medical Journal

Article Title: Suppression of METTL3 expression attenuated matrix stiffness-induced vaginal fibroblast-to-myofibroblast differentiation and abnormal modulation of the extracellular matrix in pelvic organ prolapse

doi: 10.1097/cm9.0000000000003409

Figure Lengend Snippet: Figure 2: (A–E) Representative gene expression of vaginal fibroblasts cultured on PA gels with variable stiffness (0.2 kPa vs. 12 kPa). WB analysis of protein expression and RT–qPCR analysis of the mRNA expression of α-SMA (A), COL1A1 (B), TIMP1 (C), TIMP2 (D), COL1A1/COL3A1 (E). n = 5; (F) EdU assays comparing vaginal fibroblast proliferation between 0.2 kPa and 12 kPa. (G) Representative images (IF) showing the effect of matrix stiffness on α-SMA, a marker of fibroblast-to-myofibroblast differentiation. GAPDH was used as an internal control. Scale bar = 100 µm. *P <0.05, †P <0.01, ‡P <0.001. α-SMA: α-smooth muscle actin; COL1A1: Collagen Type 1 Alpha 1; COL3A1: Collagen Type 3 Alpha 1; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: Glyceraldehyde-phosphate dehydroge nase; IF: Immunofluorescence; PA: Polyacrylamide; RT–qPCR: Real-time polymerase chain reaction; TIMP1: Tissue inhibitor of matrix metalloproteinase 1; TIMP2: Tissue inhibitor of matrix metalloproteinase 2; WB: Western blot.

Article Snippet: The membranes were blocked for 30 min and incubated with the following antibodies overnight at 4°C: anti-METTL3 (1:1000; rabbit polyclonal antibody, ABclonal; Wuhan, China); anti-α-smooth muscle actin (α-SMA) (1:1000; rabbit monoclonal antibody, ABclonal); anti-collagen Type I Alpha 1 (COL1A1) (1:800; rabbit polyclonal antibody, ABclonal); anti-collagen Type 3 Alpha 1 (COL3A1) (1:800; rabbit monoclonal antibody, ABclonal); anti-tissue inhibitor of matrix metalloproteinase 1 (TIMP1) (1:1000; rabbit polyclonal antibody, ABclonal); anti-tissue inhibitor of matrix metalloproteinase 2 (TIMP2) (1:1000; Proteintech, rabbit polyclonal antibody, Wuhan, China); anti-glyceraldehyde-phosphate dehydrogenase (GAPDH) (1:1000; rabbit monoclonal antibody, ABclonal).

Techniques: Gene Expression, Cell Culture, Expressing, Quantitative RT-PCR, Marker, Control, Immunofluorescence, Real-time Polymerase Chain Reaction, Western Blot