Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A glucuronated flavone TMMG spatially targets chondrocytes to alleviate cartilage degeneration through negative regulation of IL-1β.

doi: 10.1016/j.biopha.2023.114809

Figure Lengend Snippet: Fig. 8. Effect of TMMG administration on Elevated Serum Markers in ACLT Animals. In the serum, (A) pro-inflammatory cytokine IL-1β, (B) Carboxy-terminal telopeptides CTX-II, and (C) COMP (Cartilage oligomeric matrix protein) were elevated in the ACLT-induced osteoarthritis model. These serum bio-markers are diagnostic markers of osteoarthritis. TMMG regulated the levels of these markers in the ACLT group. All values are expressed as mean ± S.E.M. (n = 4–6/group); *P < 0.05, * *P < 0.01, and * **P < 0.001 vs. control group, and #P < 0.05, ##P < 0.01, and ###P < 0.001 vs. ACLT group.

Article Snippet: The TUNEL assay kit (C10618) was purchased from Molecular Probes Life Technologies, Rat Type II collagen Cterminal cross-linked peptide (CTX-II, E-EL-R2554) and cartilage oligomeric matrix protein (COMP, E-EL-R0159), ELISA kits were purchased from Elabscience (Elabsciences Biotechnology Co., Ltd., Hubei province, China).

Techniques: Diagnostic Assay, Control

Journal: Scientific Reports

Article Title: Lubricin binds cartilage proteins, cartilage oligomeric matrix protein, fibronectin and collagen II at the cartilage surface

doi: 10.1038/s41598-017-13558-y

Figure Lengend Snippet: Immunohistochemical co-localisation of lubricin and COMP. Dual antibody immunofluorescence on OA cartilage biopsy cryosections with anti-lubricin (P3-118) and anti-COMP (mAb 16F12) antibodies. Section shows an area of undisrupted cartilage surface (left side) and an area more severely affected by OA including a tear (right side). ( a ) Haemotoxylin and esosin staining. ( b ) DAPI staining for nuclei of chondrocytes. ( c ) COMP (green) was constantly distributed over the section with greater intensity on the superficial zone. ( d ) Lubricin (red) was present on the superficial zone of the cartilage and into the superficial zone in the area of the tissue with OA degradation at the surface. ( e ) Cartilage surface alone including COMP antibody staining, lubricin antibody staining and merged image showing co-localisation of COMP and lubricin on cartilage surface. Negative controls of dual antibody immunofluorescence analysis. The specificity of the staining was verified using matched isotype negative controls or control serum at the same concentration as the primary antibodies. ( f ) Negative control showing DAPI staining. ( g ) Negative control for the anti-COMP antibody. ( h ) Negative control for the anti-lubricin antibody.

Article Snippet: Negative controls (normal serum or isotype matched negative controls) or primary antibodies (2 µg/ml), mouse anti-human COMP (clone 16F12, Biovendor R&D) and polyclonal rabbit anti-lubricin (P3-118, Thermo Scientific) were incubated separately.

Techniques: Immunohistochemical staining, Immunofluorescence, Staining, Concentration Assay, Negative Control

Journal: Scientific Reports

Article Title: Lubricin binds cartilage proteins, cartilage oligomeric matrix protein, fibronectin and collagen II at the cartilage surface

doi: 10.1038/s41598-017-13558-y

Figure Lengend Snippet: Immunohistochemical localisation of lubricin in WT and COMP KO mice paw joint cartilage tissue. ( a ) WT mouse tissue stained with rabbit anti-lubricin (P3-118) visualised with Rhodamine Red-X shows a diffuse lubricin layer at the cartilage surface. ( b ) COMP KO mouse tissue stained with rabbit anti-lubricin (P3-118) visualised with Rhodamine Red-X showing a discrete, distinct layer of lubricin at the cartilage surface. ( c ) WT mouse tissue stained with rabbit anti-lubricin (P3-118) visualised with Rhodamine Red-X also showing DAPI staining. ( d ) COMP KO mouse tissue stained with rabbit anti-lubricin (P3-118) visualised with Rhodamine Red-X also showing DAPI staining. ( e ) Negative control of WT mouse paw tissue performed by omitting primary antibody.

Article Snippet: Negative controls (normal serum or isotype matched negative controls) or primary antibodies (2 µg/ml), mouse anti-human COMP (clone 16F12, Biovendor R&D) and polyclonal rabbit anti-lubricin (P3-118, Thermo Scientific) were incubated separately.

Techniques: Immunohistochemical staining, Staining, Negative Control

Journal: Scientific Reports

Article Title: Lubricin binds cartilage proteins, cartilage oligomeric matrix protein, fibronectin and collagen II at the cartilage surface

doi: 10.1038/s41598-017-13558-y

Figure Lengend Snippet: In situ PLA of lubricin with possible cartilage binding partners. In situ PLA was performed on OA cartilage biopsy cryosections with the following pairs. In situ PLA uses two primary antibodies targeting proteins of interest and secondary antibodies conjugated to DNA oligonucleotides (PLA probes). If the two proteins of interest are in a complex the two PLA probes will be in close enough proximity to bind and facilitate formation of a circular DNA molecule using two additional DNA oligonucleotides which then undergo rolling circle amplification to enhance the signal. Fluorescently (red) labelled complementary DNA is then added in order to visualise the DNA. ( a ) Lubricin (PA3-118) and COMP (16F12). Bound lubricin and COMP (red) were identified at the cartilage surface. A second layer was also apparent below the surface. ( b ) Lubricin (5C11) and fibronectin (ab32419). Bound lubricin and fibronectin (red) were predominately identified at the cartilage surface. Only when the PLA signal was thinner at the surface was signal observed just below the surface. ( c ) Lubricin (5C11) and collagen II (sc-7763). Bound lubricin and collagen II (red) were identified only at the surface with no PLA signal further into the tissue. PLA signal are red, DAPI staining in blue. For the 3 antibody pairs, 5 separate antibodies were used as anti-lubricin 5CII was used for 2 pairs. Negative controls were performed by omitting each primary antibody for all of the five antibodies used. ( d ) lubricin negative control (PA3-118), ( e ) COMP negative control (16F12), ( f ) lubricin negative control (5CII), ( g ) fibronectin negative control (ab32419) and ( h ) collagen II negative control (sc-7763).

Article Snippet: Negative controls (normal serum or isotype matched negative controls) or primary antibodies (2 µg/ml), mouse anti-human COMP (clone 16F12, Biovendor R&D) and polyclonal rabbit anti-lubricin (P3-118, Thermo Scientific) were incubated separately.

Techniques: In Situ, Binding Assay, Amplification, Staining, Negative Control

Journal: Scientific Reports

Article Title: Lubricin binds cartilage proteins, cartilage oligomeric matrix protein, fibronectin and collagen II at the cartilage surface

doi: 10.1038/s41598-017-13558-y

Figure Lengend Snippet: Illustration of the cartilage surface showing protein interactions. Lubricin is blue with glycosylation shown in light blue, COMP is red shown as the pentameric structure, fibronectin is green shown as the dimeric structure and collagen II is purple shown as collagen II fibrils. The SF is shown in light straw, the cartilage in yellow and the cartilage surface in pink. COMP, collagen II and fibronectin adhere lubricin, primarily by the termini of lubricin, to the cartilage surface creating an exposed lubricin layer. The ECM proteins also bind to each other and are shown in the cartilage. Lubricin is also found in the SF.

Article Snippet: Negative controls (normal serum or isotype matched negative controls) or primary antibodies (2 µg/ml), mouse anti-human COMP (clone 16F12, Biovendor R&D) and polyclonal rabbit anti-lubricin (P3-118, Thermo Scientific) were incubated separately.

Techniques: