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

Journal: Scientific Reports

Article Title: Glycation damage to organelles and their DNA increases during maize seedling development

doi: 10.1038/s41598-022-06454-7

Figure Lengend Snippet: AGE in maize mitochondria and plastids. The amount of AGE in proteins extracted from isolated mitochondria and plastids was measured using anti-AGE antibodies with a competitive ELISA ( A – D ) and slot blot ( E – L ) assays. For the ELISA assay the levels of AGE in total mitochondrial and plastid proteins were measured as µg/mL. Representative images of slot blot assays are shown in ( I–L ) and the relative AGE in protein was determined, where the image intensity was set to 1 for stalk lower in ( E , G ) and for dark-grown leaves in ( F , H ). Uppermost panels of ( I – L ) show AGE proteins, middle panels show the total protein (TP) levels in samples, and lowermost panels show mitochondrial (COX2)- and plastid (RbcL)-specific proteins (COX2 in panels I and J; RbcL in panels K and L). Equal amounts of protein extracted from the different tissues were used in these assays. Data here and in Figs. , , , are represented as mean ± SEM. Statistically significant differences were measured using ANOVA statistic test with post hoc analysis using Tukey’s HSD and are shown as asterisks, where * P -value ≤ 0.05, ** P -value ≤ 0.01, *** P -value ≤ 0.001 are indicated on respective graphs.

Article Snippet: Additionally, to ensure equal amounts of housekeeping proteins in the samples, the proteins were loaded on the membrane and probed with anti-RbcL antibodies for plastids (1:1000 dilution) (PhytoAB; cat # PHY1413S) and anti-COX2 antibodies for mitochondria (1:1000 dilution) (PhytoAB; cat # PHY0066S).

Techniques: Isolation, Competitive ELISA, Dot Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Glycation damage to organelles and their DNA increases during maize seedling development

doi: 10.1038/s41598-022-06454-7

Figure Lengend Snippet: DJ-1 protein in maize mitochondria and plastids. The amount of DJ-1 protein from isolated mitochondria and plastids was measured using anti-DJ-1 antibodies with competitive ELISA ( A – D ) and slot blot ( E – H ) assays. For ELISA, DJ-1 protein in mitochondria was measured as ng/mL. Representative images of slot blot assays are shown ( I – L ) and the relative DJ-1 protein was determined, where the image intensity was set to 1 for leaf 1 (L1) in ( E , G ) and for light-grown leaves in ( F , H ). Uppermost panels of I-L show DJ-1 proteins, middle panels show the total protein (TP) levels in samples, and lowermost panels show mitochondrial (COX2)- and plastid (RbcL)-specific proteins (COX2 in panels I and J; RbcL in panels K and L). Cross-reactivity of maize organellar DJ-1 with anti-DJ-1 antibodies (human Park7) was confirmed and is shown in Supplementary Information, Fig. S1.

Article Snippet: Additionally, to ensure equal amounts of housekeeping proteins in the samples, the proteins were loaded on the membrane and probed with anti-RbcL antibodies for plastids (1:1000 dilution) (PhytoAB; cat # PHY1413S) and anti-COX2 antibodies for mitochondria (1:1000 dilution) (PhytoAB; cat # PHY0066S).

Techniques: Isolation, Competitive ELISA, Dot Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Glycation damage to organelles and their DNA increases during maize seedling development

doi: 10.1038/s41598-022-06454-7

Figure Lengend Snippet: Methylglyoxal (MGO) in maize organelles. Mitochondria and plastids were isolated and MGO levels in nmole determined with a MGO assay kit as per manufacturer’s instructions. ( A and B ) show MGO in mitochondria; ( C and D ) show MGO in plastids.

Article Snippet: Additionally, to ensure equal amounts of housekeeping proteins in the samples, the proteins were loaded on the membrane and probed with anti-RbcL antibodies for plastids (1:1000 dilution) (PhytoAB; cat # PHY1413S) and anti-COX2 antibodies for mitochondria (1:1000 dilution) (PhytoAB; cat # PHY0066S).

Techniques: Isolation

Journal: Scientific Reports

Article Title: Glycation damage to organelles and their DNA increases during maize seedling development

doi: 10.1038/s41598-022-06454-7

Figure Lengend Snippet: AGE in DNA isolated from mitochondria and plastids. For competitive ELISA assays ( A – D ), AGE levels were determined as µg/mL. The relative levels of AGE were also determined by slot blot assays ( E – H ), and representative images are shown ( I – L ) where the image intensity was set to 1 for Stalk lower in ( E , G ) and for dark-grown leaves in ( F , H ). Equal amounts of mtDNA extracted from the different tissues were used in these assays. Lower panels of ( I – L ) show the equal amount of orgDNA as determined by slot blot using mitochondrial- and plastid-specific probes (cox2 and rbcL, respectively).

Article Snippet: Additionally, to ensure equal amounts of housekeeping proteins in the samples, the proteins were loaded on the membrane and probed with anti-RbcL antibodies for plastids (1:1000 dilution) (PhytoAB; cat # PHY1413S) and anti-COX2 antibodies for mitochondria (1:1000 dilution) (PhytoAB; cat # PHY0066S).

Techniques: Isolation, Competitive ELISA, Dot Blot

Journal: Scientific Reports

Article Title: Glycation damage to organelles and their DNA increases during maize seedling development

doi: 10.1038/s41598-022-06454-7

Figure Lengend Snippet: Summary of results from the assays.

Article Snippet: Additionally, to ensure equal amounts of housekeeping proteins in the samples, the proteins were loaded on the membrane and probed with anti-RbcL antibodies for plastids (1:1000 dilution) (PhytoAB; cat # PHY1413S) and anti-COX2 antibodies for mitochondria (1:1000 dilution) (PhytoAB; cat # PHY0066S).

Techniques: