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: 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

Journal: Breast Cancer Research : BCR

Article Title: BRCA1 mutation influences progesterone response in human benign mammary organoids

doi: 10.1186/s13058-019-1214-0

Figure Lengend Snippet: Confirmation and validation of ECM genes . a Confirmation of ECM genes was done by real-time PCR analysis of the samples used in the RNA-seq analysis. The data are fold changes of hormone treatment (dotted red line) and represent the mean ± SEM from three technical replicates (* p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001). b Validation of ECM genes was done by real-time PCR using new patient-derived individual BRCA1 mut and non-carrier organoids treated with E2+P4 and E2+P4+TPA. The data are fold changes of hormone treatment (dotted red line) and represent the mean ± SEM from three technical replicates (* p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001. c IHC staining of ECM proteins MMP10, COL1A1, COL3A1, and FN1 were performed in non-carrier and BRCA1 mut organoids. Scale bar, 100 μm

Article Snippet: The primary antibodies used were ER (1:200, Santa Cruz SC71064), PR (1:200, DAKO, M3568), pan cytokeratin (panCK; 1:200, Abcam, ab7753), Cytokeratin 18 (CK18; 1:200, Abcam, 93741), vimentin (Vim; 1:200, Abcam, 92547), α smooth muscle actin (αSMA; 1:200, Novus Biologicals, 600531), Cytokeratin 5/6 (CK5/6; 1:200, Agilent Technologies, M723729-2), COL3A1 (Atlas antibodies, 1:200, HPA007583), COL1A1 (LS Bio, 1:400, LS-C343921), MMP10 (LS Bio, 1:400, LS-C118518), MMP1(LS Bio, 1:400, LS-B8645), Ki67 (Dako, 1:200, GA62661-2), E-cadherin (BD Biosciences, 1:200, 610181), ITGB4 (LS Bio, 1:200, LS-B3778), and ALDH1A1 isoform (LSBio, 1:400, LS-B10149).

Techniques: Biomarker Discovery, Real-time Polymerase Chain Reaction, RNA Sequencing, Derivative Assay, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Intermittent Fasting Inhibits High-Fat Diet–Induced Atherosclerosis by Ameliorating Hypercholesterolemia and Reducing Monocyte Chemoattraction

doi: 10.3389/fphar.2021.719750

Figure Lengend Snippet: Sequences of primers for qRT-PCR.

Article Snippet: Mouse anti–smooth muscle α-actin (SMA) monoclonal antibody and rabbit anti-collagen type III alpha 1 (COL3A1) polyclonal antibody were purchased from Affinity Biosciences (Cincinnati, OH, United States).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Intermittent Fasting Inhibits High-Fat Diet–Induced Atherosclerosis by Ameliorating Hypercholesterolemia and Reducing Monocyte Chemoattraction

doi: 10.3389/fphar.2021.719750

Figure Lengend Snippet: Intermittent fasting increases collagen content via inducing COL1A1 and COL3A1 expression. (A,B) The aortic root cross sections were used to determine collagen content by Picrosirius red staining with quantitative analysis of collagen positive areas. * p < 0.05 ( n = 6). (C,D) The expression of COL1A1, COL3A1, MMP2, and MMP9 in plaques were determined by immunofluorescent staining with the quantification of positive staining areas. * p < 0.05, ns: not significant ( n = 6). E : LDLR−/− mice were divided into the control or IF group ( n = 6/group). After 2 weeks HFD feeding ad libitum or IF treatment, the mice aorta samples were collected. The expression of COL1A1, COL3A1, MMP2, and MMP9 were determined by qRT-PCR. * p < 0.05, ns: not significant ( n = 6).

Article Snippet: Mouse anti–smooth muscle α-actin (SMA) monoclonal antibody and rabbit anti-collagen type III alpha 1 (COL3A1) polyclonal antibody were purchased from Affinity Biosciences (Cincinnati, OH, United States).

Techniques: Expressing, Staining, Control, Quantitative RT-PCR