Journal: Journal of Biological Chemistry

Article Title: Modulation of matrix metabolism by ATP-citrate lyase in articular chondrocytes

doi: 10.1074/jbc.ra118.002261

Figure Lengend Snippet: Figure 5. Inhibition of ACLY by HCA attenuated IL-1β-induced augmentation of acetylation H3K9 and H3K27 both globally and specifically in the iNOS, MMP3 and MMP13 promoters.

Article Snippet: Assays of nitric oxide (NO), MMP3, and MMP13 release, and SOX9 acetylation, distribution, and expression Conditioned media were tested for NO generation by Griess reaction assay to quantify nitrite, and MMP3 and MMP13 release measured using Total MMP3 and MMP13 Quantikine ELISA Kits (R&D Systems).

Techniques: Inhibition

Journal: Cell Death and Differentiation

Article Title: Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis

doi: 10.1038/s41418-020-0505-4

Figure Lengend Snippet: a Western blot analysis and semiquantitative evaluation of DLL4, VEGFA, and CD31 expression in PDX mice tissues by densitometry analysis of protein bands reveals a downregulation of DLL4, VEGFA, and CD31 protein expression in PDX mice treated with GSI. The bands were measured compared with the housekeeping GAPDH protein band, for each tissue. Average value of DLL4, VEGFA, and CD31 expression levels among all mouse treated with LY3039478 or vehicle is reported in the graph. P value showed versus vehicle treatment. Tissues PDX mice n = 10 for vehicle treatment in gray, n = 10 for LY3039478 treatment in black. b Representative images with immunofluorescence staining show DLL4 and CD31 downregulation in representative images of PDX tissues treated with LY30349478. DLL4 (green) and CD31 (red) and overlapping staining (yellow) were immunolocalized in PDX tissues. The yellow arrows highlight the detail of the co-localization of DLL4 and CD31 in PDX tissues (#4, #14, #24) not treated with LY339478. DAPI, 4′,6‐diamidino‐2‐phenylindole. c Immunofluorescence staining with MMP13 in red and nucleus in DAPI shown a significantly reduction of MMP13 in iCCA PDX tissues treated with LY3039478. Magnifications: ×20; inset ×60. d Representative images demonstrate a significant ( P < 0.001) destruction of the network created by the HUVECs following the treatment with LY3039478 (1 µM). The concomitant administration of MMP13 counteracts significantly ( P < 0.01) drug effectiveness.

Article Snippet: The recombinant protein MMP13 (R&D System, Minneapolis, MN) was used at 50 ng/ml.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Cell Death and Differentiation

Article Title: Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis

doi: 10.1038/s41418-020-0505-4

Figure Lengend Snippet: a Analysis of 31 primary tumors from iCCA patients and matched surrounding normal liver tissues downloaded from the GEO database (GSE107943). Mean expression data were expressed in RPKM (Reads Per Kilobase Million). *** P < 0.001 calculated with Student’s t test. b NOTCH1 gene and its pro-angiogenic targets are overexpressed in human intrahepatic cholangiocarcinoma (iCCA). Levels of NOTCH1, DLL4, VEGFA, and MMP13 mRNA were significantly more elevated in iCCA ( n = 42) than corresponding nontumorous surrounding livers (SL; n = 42), as detected by quantitative reverse-transcription PCR. Number target (NT) = 2 −ΔCt , wherein ΔCt value of each sample was calculated by subtracting the average Ct value of the gene of interest from the average Ct value of the β-actin gene. Mann–Whitney test: vs SL, P < 0.0001. c Expression of the NOTCH1 gene correlates with mRNA levels of putative target genes (HES1, DLL4, VEGFA, and MMP13) in a collection of human intrahepatic cholangiocarcinoma (CCA) samples ( n = 42). Linear regression analysis was used. d Representative expression patterns of CK19, NOTCH1, HES1, DDL4, and MMP13 in human intrahepatic cholangiocarcinoma (iCCA) as detected by immunohistochemistry. Upper panels: CCA case (CCA1) showing strong, concomitant immunoreactivity for NOTCH1, HES1, DDL4, and MMP13. Lower panels: CCA specimens (CCA2) exhibiting low levels of NOTCH1, HES1, DDL4, and MMP13. As expected, both iCCA display robust immunolabeling for CK19 (a biliary marker). Magnification: ×200; scale bar = 100 μm. H&E hematoxylin and eosin staining.

Article Snippet: The recombinant protein MMP13 (R&D System, Minneapolis, MN) was used at 50 ng/ml.

Techniques: Expressing, Reverse Transcription, MANN-WHITNEY, Immunohistochemistry, Immunolabeling, Marker, Staining

Journal: Cell Death and Differentiation

Article Title: Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis

doi: 10.1038/s41418-020-0505-4

Figure Lengend Snippet: Levels of tumor microvessel density (MVD) correlate with mRNA expression of NOTCH1 ( a ), HES1 ( b ), DLL4 ( c ), and MMP13 ( e ), but not with those of VEGFA ( d ), in a collection of human intrahepatic cholangiocarcinoma (iCCA) samples ( n = 42). Linear regression analysis was used. f Representative examples of human iCCA specimens with high and low MVD.

Article Snippet: The recombinant protein MMP13 (R&D System, Minneapolis, MN) was used at 50 ng/ml.

Techniques: Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secoisolariciresinol diglucoside Ameliorates Osteoarthritis via Nuclear factor-erythroid 2-related factor-2/ nuclear factor kappa B Pathway: In vitro and in vivo experiments.

doi: 10.1016/j.biopha.2023.114964

Figure Lengend Snippet: Fig. 3. The effect of SDG on IL-1β-stimulated inflammation and ECM degradation in SW1353. (A)The protein expressions of COL2A1, ADAMTS5, SOX9, MMP13, INOS and COX2 in SW1353 were evaluated by western blotting. (B)The ADAMTS5 were detected by fluorescence microscopy, combined with nuclear DAPI staining (scale bar: 200 µm). (C)The COL2A1 were detected by fluorescence microscopy, combined with nuclear DAPI staining (scale bar: 200 µm).The values are mean ± SD for 3 separate experiments. *p < 0.05, **p < 0.01 and ***p < 0.001. SDG, secoisolariciresinol diglucoside; ECM, extracellular matrix; COL2A1, collagen II; ADAMTS5, a disintegrin and metalloproteinase with thrombospondin motifs 5; SOX9, SRY-related high-mobility-group-box gene 9; MMP13, matrix metal loproteinases 13; INOS, inducible nitric oxide synthase; COX2, cyclooxygenase-2.

Article Snippet: Primary antibodies against Actin, COL2A1, ADAMTS5, SOX9, MMP13, INOS, COX2, Nrf2, HO-1 and Z. Zhang et al. Biomedicine & Pharmacotherapy 164 (2023) 114964 IκBαwere bought from Boster (Wuhan, China).

Techniques: Western Blot, Fluorescence, Microscopy, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secoisolariciresinol diglucoside Ameliorates Osteoarthritis via Nuclear factor-erythroid 2-related factor-2/ nuclear factor kappa B Pathway: In vitro and in vivo experiments.

doi: 10.1016/j.biopha.2023.114964

Figure Lengend Snippet: Fig. 4. The effect of SDG on IL-1β-stimulated inflammation and ECM degradation in mouse primary chondrocytes. The protein expressions of COL2A1, ADAMTS5, SOX9, MMP13, INOS and COX2 in mouse primary chondrocytes were assessed by western blotting. The values are mean ± SD for 3 separate experiments. *p < 0.05, **p < 0.01 and ***p < 0.001. SDG, secoisolariciresinol diglucoside; ECM, extracellular matrix; IL-1β, interleukin-1β; COL2A1, collagen II; ADAMTS5, a disintegrin and metalloproteinase with thrombospondin motifs 5; SOX9, SRY-related high-mobility-group-box gene 9; MMP13, matrix metalloproteinases 13; INOS, inducible nitric oxide synthase; COX2, cyclooxygenase-2.

Article Snippet: Primary antibodies against Actin, COL2A1, ADAMTS5, SOX9, MMP13, INOS, COX2, Nrf2, HO-1 and Z. Zhang et al. Biomedicine & Pharmacotherapy 164 (2023) 114964 IκBαwere bought from Boster (Wuhan, China).

Techniques: Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secoisolariciresinol diglucoside Ameliorates Osteoarthritis via Nuclear factor-erythroid 2-related factor-2/ nuclear factor kappa B Pathway: In vitro and in vivo experiments.

doi: 10.1016/j.biopha.2023.114964

Figure Lengend Snippet: Fig. 7. The protective effects of SDG in IL-1β-activated SW1353 were reversed by Nrf2-siRNA. (A)The protein expressions of Nrf2 and HO-1 were evaluated by western blotting in chondrocytes transfected with Nrf2 siRNA. (B)The protein expressions of COL2A1, ADAMTS5, SOX9, MMP13, INOS and COX2 was assessed by western blotting in chondrocytes transfected with Nrf2 siRNA. The values are mean ± SD for 3 separate experiments. *p < 0.05, **p < 0.01 and ***p < 0.001. SDG, secoisolariciresinol diglucoside; IL-1β, interleukin-1β; Nrf2, Nuclear factor-erythroid 2-related factor-2; HO-1, Heme Oxygenase-1; COL2A1, collagen II; ADAMTS5, a disintegrin and metalloproteinase with thrombospondin motifs 5; SOX9, SRY-related high-mobility-group-box gene 9; MMP13, matrix metalloproteinases 13; INOS, inducible nitric oxide synthase; COX2, cyclooxygenase-2.

Article Snippet: Primary antibodies against Actin, COL2A1, ADAMTS5, SOX9, MMP13, INOS, COX2, Nrf2, HO-1 and Z. Zhang et al. Biomedicine & Pharmacotherapy 164 (2023) 114964 IκBαwere bought from Boster (Wuhan, China).

Techniques: Western Blot, Transfection