Journal: Life Science Alliance

Article Title: Local tissue mechanics control cardiac pacemaker cell embryonic patterning

doi: 10.26508/lsa.202201799

Figure Lengend Snippet: (A) Col3A1 staining at the SAN/atria junction in representative HH18, HH30, and HH35 hearts. (B) Distribution of Col3A1, TnC, and WFA in the H35 SAN and atria. (C) Diagram of in vivo transfection strategy for mosaic calcium imaging in the intact SAN. Integrating DNA plasmids were microinjected into the pericardial space adjacent to the forming SAN at HH18 and SAN tissue was isolated for live imaging at HH30. (D) Live imaging of a memRFP/GCaMP6f-positive cell within the SAN. Data presented in were taken from cells imaged in 25 individual hearts.

Article Snippet: The following reagents were used for immunohistochemistry: MF20 (14650380; Invitrogen) (1:500); FLRT3 (PA563240; Invitrogen) (1:250); hyaluronic acid binding protein, bovine nasal cartilage, biotinylated (38591150UG; MilliporeSigma) (1:100); Wisteria floribunda lectin biotinylated (B13552; Vector Laboratories) (1:100); Col3A1 (3B2-s; DSHB) (1:250), TNC (LS-C39574-50; LSBio) (1:500); Nav1.5 (LS-C30483; LSBio) (1:500), phalloidin-647 (A22287; Invitrogen) (1:40); NCX1 (LS-C73201-100; LSBio) (1:500); streptavidin-568 (S11226; Invitrogen) (1:500); DAPI (62248; Thermo Fisher Scientific) (1:1,000); IgM (heavy chain) cross-adsorbed goat anti-mouse, Alexa Fluor 555 (A21426; Invitrogen) (1:500); IgG1 cross-adsorbed goat anti-mouse, Alexa Fluor 647 (A21240; Invitrogen) (1:500); IgG2b cross-adsorbed goat anti-mouse, Alexa Fluor 488 (A21141; Invitrogen) (1:500); IgG (H+L) cross-adsorbed goat anti-rabbit, Alexa Fluor 568 (A11011; Invitrogen) (1:500); goat anti-mouse IgG2b cross-adsorbed secondary antibody, Alexa Fluor 647 (A21242; Invitrogen) (1:500); goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody, Alexa Fluor 488 (A11008; Invitrogen) (1:500); goat anti-mouse IgM mu chain Alexa Fluor 488 (ab150121; Abcam) (1:500).

Techniques: Staining, In Vivo, Transfection, Imaging, Isolation

Journal: STAR Protocols

Article Title: Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing

doi: 10.1016/j.xpro.2025.104296

Figure Lengend Snippet: 3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Article Snippet: Collagen III antibody, anti-human, PE, REAdye_lease , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-357 RRID: AB_2905171.

Techniques: Imaging, Comparison, Selection, Staining

Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 3. TXNIP promoted post-MI collagen deposition (A,B) Representative picrosirius red staining images of heart sections from each group at 28 days after treatment (A) under a light microscope and (B) under a polarized light microscope. Collagen I showed red/orange birefringence, while Collagen III showed green/whitish birefringence. (C,E,G) Representative immunohistochemical staining images of heart sections from each group at 28 days after treatment. (C) Collagen I deposition. (E) Collagen III deposition, and (G) ACTA2 deposition. (D) Statistical analysis of Collagen I deposition. Data are shown as the mean±SEM, n=3. **P<0.01. (F) Statistical analysis of Collagen III deposition. Data are shown as the mean± SEM, n=3. **P<0.01. (H) Statistical analysis of ACTA2 deposition. Data are shown as the mean±SEM, n=3. *P<0.05, **P<0.01.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques: Staining, Light Microscopy, Immunohistochemical staining

Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 4. TXNIP increased post-MI collagen deposition (A) Representative images of the protein levels of Collagen III, Collagen I, ACTA2, and CTGF. (B‒E) Statistical analyses of the protein levels of Collagen I, Collagen III, ACTA2, and CTGF. Data are shown as the mean±SEM, n=3‒5. *P<0.05, **P<0.01; ns, not significant.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques:

Journal: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Histological evaluation of the kidney in SD and SDT fatty rats. Immunohistological staining of kidney tissues using antibodies against CD68 (a, b), α -SMA (c, d), type I collagen (e, f), and type III collagen (g, h). Histological PAS staining showing focal glomerular sclerosis (i) and semiquantitative assessment of focal glomerular sclerosis (j). Original magnification: ×100. Values are presented as the mean ± SEM. ∗ p < 0.05 vs. SD rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Staining

Journal: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Relationships between renal morphological change and muscle strength, and weights of the soleus and extensor digitorum longus (EDL) muscles in 24-week-old SD and SDT fatty rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Muscles, Expressing

Journal: International Journal of Molecular Medicine

Article Title: Role of transforming growth factor β-1 in the pathogenesis of pelvic organ prolapse: A potential therapeutic target

doi: 10.3892/ijmm.2017.3042

Figure Lengend Snippet: Sequences of primers used for RT-qPCR.

Article Snippet: The antibodies used were as follows: COL1A1 antibody (diluted at 1/100; BA0325; Boster Inc., Wuhan, China), COL3A1 antibody (diluted at 1/150; BA0326; Boster Inc.), elastin antibody (diluted at 1/100; ab23747; Abcam, Cambridge, UK), MMP-2 antibody (diluted at 1/75; BA3716; Boster Inc.), MMP-9 antibody (diluted at 1/100; BA0573; Boster Inc.), TIMP-2 antibody (diluted at 1/100; sc-5539; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) TGF-β1 antibody (diluted at 1/100; ab92486; Abcam).

Techniques: Sequencing

Journal: International Journal of Molecular Medicine

Article Title: Role of transforming growth factor β-1 in the pathogenesis of pelvic organ prolapse: A potential therapeutic target

doi: 10.3892/ijmm.2017.3042

Figure Lengend Snippet: Expression levels of collagen fibers and elastin in USL tissue from the control group and the POP group. (A) Upper panel, by Masson's trichrome staining, collagen fibers are stained blue, smooth muscle and cytoplasm are stained red, cell nuclei are stained black; second panel, immunohistochemical staining of elastin fiber, strong positive in the control group, moderately positive in the POP group; third panel, statistical analysis based on the IOD value and mean density captured by Image-Pro plus 6.0 software. (B) The mRNA expression levels of COL1A1, COL3A1 and elastin detected by RT-qPCR. Data are presented as the means ±standard deviation for at least 3 independent experiments. n=30; * P<0.05 vs. the control group, ** P<0.01 vs. the control group. USL, uterosacral ligament; POP, pelvic organ prolapse; COL, collagen; IOD, integrated optical density.

Article Snippet: The antibodies used were as follows: COL1A1 antibody (diluted at 1/100; BA0325; Boster Inc., Wuhan, China), COL3A1 antibody (diluted at 1/150; BA0326; Boster Inc.), elastin antibody (diluted at 1/100; ab23747; Abcam, Cambridge, UK), MMP-2 antibody (diluted at 1/75; BA3716; Boster Inc.), MMP-9 antibody (diluted at 1/100; BA0573; Boster Inc.), TIMP-2 antibody (diluted at 1/100; sc-5539; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) TGF-β1 antibody (diluted at 1/100; ab92486; Abcam).

Techniques: Expressing, Control, Staining, Immunohistochemical staining, Software, Quantitative RT-PCR, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Role of transforming growth factor β-1 in the pathogenesis of pelvic organ prolapse: A potential therapeutic target

doi: 10.3892/ijmm.2017.3042

Figure Lengend Snippet: Effect of TGF-β1 on cell viability and ECM metabolism in the control or hUSLFs subjected to CMS. In the study groups, hUSLFs were incubated with recombinant human TGF- 1 at concentrations of 0, 5, or 10 ng/ml for 24 h, then exposed to CMS at a strain of 5,333 μ ε for 4 h. The normal control group was not treated with TGF-β1 or CMS. (A) Cell viability was detected using the Cell Counting kit-8 (CCK-8) method. Expression levels of COL1A1, COL3A1, elastin, TIMP-2, MMP-9 and MMP-2 were analyzed by (B) western blot analysis, (C) gelatin zymography, or (D) RT-qPCR. Data represent the means ± standard deviation for at least 3 independent experiments; n=3; * P<0.05 vs. the control group; # P<0.05 vs. the CMS group. TGF, transforming growth factor ECM, extracellular matrix; CMS, cyclic mechanical stretching; hUSLF, human USL fibroblast; COL, collagen; TIMP, tissue inhibitor of MMP; MMP, matrix metallloproteinase.

Article Snippet: The antibodies used were as follows: COL1A1 antibody (diluted at 1/100; BA0325; Boster Inc., Wuhan, China), COL3A1 antibody (diluted at 1/150; BA0326; Boster Inc.), elastin antibody (diluted at 1/100; ab23747; Abcam, Cambridge, UK), MMP-2 antibody (diluted at 1/75; BA3716; Boster Inc.), MMP-9 antibody (diluted at 1/100; BA0573; Boster Inc.), TIMP-2 antibody (diluted at 1/100; sc-5539; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) TGF-β1 antibody (diluted at 1/100; ab92486; Abcam).

Techniques: Control, Incubation, Recombinant, Cell Counting, CCK-8 Assay, Expressing, Western Blot, Zymography, Quantitative RT-PCR, Standard Deviation