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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
Immunofluorescence Labeling, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
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mitotracker green immunofluorescent double labelling  (Beyotime)
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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
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vectafluor duet immunofluorescence double labeling kit  (Vector Laboratories)
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Vector Laboratories vectafluor duet immunofluorescence double labeling kit
Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
Vectafluor Duet Immunofluorescence Double Labeling Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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immunofluorescent labeling  (Nikon)
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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) <t>Immunofluorescence</t> of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.
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Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) Immunofluorescence of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.

Journal: Advanced Science

Article Title: Osteoclast‐Derived SLIT3 Mediates Osteoarthritis Pain and Degenerative Changes

doi: 10.1002/advs.202517545

Figure Lengend Snippet: Pain in TRAP‐Cre; Slit3 −/− mice is reduced. a) Experimental design for TRAP‐Cre transgenic mice injected with AAV9‐Slit3‐RNAi. b) Immunofluorescence of SLIT3 (red) in condyles. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) Representative results of OFT and EPM. e,f) Quantitative analysis of Von Frey experiment (e), OFT and EPM (f). n = 6. Statistical analyses were performed using Student's t ‐test in (c), and one‐way ANOVA with Holm–Šidák multiple comparison tests in (e,f). ** p < 0.01. *** p < 0.001. **** p < 0.0001. ns : no significance.

Article Snippet: To evaluate the axonal morphology of the TGs in the different groups (SLIT3 protein, RPD353Mu01, Cloud‐clone, USA; Neutralizing antibodies against SLIT3, AF3629, R&D Systems, USA), immunofluorescence labeling (β3‐tubulin, 1:300, 2144, Cell Signaling Technology) and crystal violet staining ( n = 6) were conducted.

Techniques: Transgenic Assay, Injection, Immunofluorescence, Comparison

Immunofluorescence imaging and analysis of sensory nerve distribution in the condyle of Slit3 ‐knockdown mice. a) Representative images of immunofluorescence staining for PGP9.5 (red), CGRP (green) and DAPI (blue) along the TMJ osteochondral junction. Scale bars: 10 µm. b) Quantitative analysis of the density of PGP9.5 + nerve fibers and CGRP + nerve fibers in the TMJ osteochondral junction. n = 6. c) Schematic diagram of anterograde tracing. d) Representative images of the condylar sensory nerve. EGFP (rAAV‐CAG‐EGFP‐WPRE‐hGH pA, green), DAPI (blue). Scale bars: 10 µm. e) Quantitative analysis of the density of sensory nerve fibers. n = 6. The white dashed line represents the boundary between bone and cartilage. Statistical analyses were performed using one‐way ANOVA with Holm–Šidák multiple comparison tests. **** p < 0.0001. ns : no significance.

Journal: Advanced Science

Article Title: Osteoclast‐Derived SLIT3 Mediates Osteoarthritis Pain and Degenerative Changes

doi: 10.1002/advs.202517545

Figure Lengend Snippet: Immunofluorescence imaging and analysis of sensory nerve distribution in the condyle of Slit3 ‐knockdown mice. a) Representative images of immunofluorescence staining for PGP9.5 (red), CGRP (green) and DAPI (blue) along the TMJ osteochondral junction. Scale bars: 10 µm. b) Quantitative analysis of the density of PGP9.5 + nerve fibers and CGRP + nerve fibers in the TMJ osteochondral junction. n = 6. c) Schematic diagram of anterograde tracing. d) Representative images of the condylar sensory nerve. EGFP (rAAV‐CAG‐EGFP‐WPRE‐hGH pA, green), DAPI (blue). Scale bars: 10 µm. e) Quantitative analysis of the density of sensory nerve fibers. n = 6. The white dashed line represents the boundary between bone and cartilage. Statistical analyses were performed using one‐way ANOVA with Holm–Šidák multiple comparison tests. **** p < 0.0001. ns : no significance.

Article Snippet: To evaluate the axonal morphology of the TGs in the different groups (SLIT3 protein, RPD353Mu01, Cloud‐clone, USA; Neutralizing antibodies against SLIT3, AF3629, R&D Systems, USA), immunofluorescence labeling (β3‐tubulin, 1:300, 2144, Cell Signaling Technology) and crystal violet staining ( n = 6) were conducted.

Techniques: Immunofluorescence, Imaging, Knockdown, Staining, Anterograde Tracing, Comparison

Slit3 is specifically knocked out in osteoclasts of Slit3 ‐CKO mice. a) Schematic diagram of Slit3 ‐CKO mice. b) Immunofluorescence of SLIT3 (green) and CD51/61 (red) in subchondral bone. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) qRT‐PCR analysis of the gene expression of Slit3 in subchondral bone in the two groups. n = 3. e,f) Western blot and its quantitative analysis, g) ELISA, and h,i) immunohistochemical staining and quantitative analysis of SLIT3 expression in subchondral bone in the WT and Slit3 ‐CKO groups. n = 3. Scale bars: 70 µm. Statistical analyses were performed using Student's t ‐test. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p <0.0001. ns : no significance.

Journal: Advanced Science

Article Title: Osteoclast‐Derived SLIT3 Mediates Osteoarthritis Pain and Degenerative Changes

doi: 10.1002/advs.202517545

Figure Lengend Snippet: Slit3 is specifically knocked out in osteoclasts of Slit3 ‐CKO mice. a) Schematic diagram of Slit3 ‐CKO mice. b) Immunofluorescence of SLIT3 (green) and CD51/61 (red) in subchondral bone. The white dashed line represents the boundary between subchondral bone and cartilage. Scale bars:10 µm. c) Quantitative analysis in panel (b). n = 6. d) qRT‐PCR analysis of the gene expression of Slit3 in subchondral bone in the two groups. n = 3. e,f) Western blot and its quantitative analysis, g) ELISA, and h,i) immunohistochemical staining and quantitative analysis of SLIT3 expression in subchondral bone in the WT and Slit3 ‐CKO groups. n = 3. Scale bars: 70 µm. Statistical analyses were performed using Student's t ‐test. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p <0.0001. ns : no significance.

Article Snippet: To evaluate the axonal morphology of the TGs in the different groups (SLIT3 protein, RPD353Mu01, Cloud‐clone, USA; Neutralizing antibodies against SLIT3, AF3629, R&D Systems, USA), immunofluorescence labeling (β3‐tubulin, 1:300, 2144, Cell Signaling Technology) and crystal violet staining ( n = 6) were conducted.

Techniques: Immunofluorescence, Quantitative RT-PCR, Gene Expression, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing