Journal: BMC Complementary Medicine and Therapies

Article Title: Treatment of rheumatoid arthritis by Rhododendron molle G. Don: the anti-inflammatory mechanism of its extract was elucidated by network analysis combined with experimental verification

doi: 10.1186/s12906-025-04982-z

Figure Lengend Snippet: Molecular docking results of 4 active metabolites of R .molle with core targets. A) Dvl1 (PDBID:6lcb). B) GSK-3β (PDBID:1o6l)

Article Snippet: The primary antibodies against β-actin (81115-1-RR), APC (19782-1-AP), Dvl1 (27384-1-AP), β-catenin (51067-2-AP), HRP-Goat Anti-Rabbit IgG (SA00001-2) were purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques:

Journal: BMC Complementary Medicine and Therapies

Article Title: Treatment of rheumatoid arthritis by Rhododendron molle G. Don: the anti-inflammatory mechanism of its extract was elucidated by network analysis combined with experimental verification

doi: 10.1186/s12906-025-04982-z

Figure Lengend Snippet: RME inhibited the expression of key proteins of Dvl1/Wnt1/β-catenin/GSK-3β signaling pathway. All data are expressed as mean ± SEM ( n = 3). *** P < 0.001compared with the CON group, # P < 0.05, ## P < 0.01, ### P < 0.001 compared with the Model group

Article Snippet: The primary antibodies against β-actin (81115-1-RR), APC (19782-1-AP), Dvl1 (27384-1-AP), β-catenin (51067-2-AP), HRP-Goat Anti-Rabbit IgG (SA00001-2) were purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Expressing

Journal: BMC Complementary Medicine and Therapies

Article Title: Treatment of rheumatoid arthritis by Rhododendron molle G. Don: the anti-inflammatory mechanism of its extract was elucidated by network analysis combined with experimental verification

doi: 10.1186/s12906-025-04982-z

Figure Lengend Snippet: The mechanism of RME inhibiting the secretion of inflammatory factors by regulating the Dvl1/Wnt1/β-catenin/GSK-3β signaling pathway in the treatment of RA. (FZD: frizzled protein, LRP: low density lipoprotein-related receptor, TCF: T cytokine, LEF: lymphocyte enhancing factor)

Article Snippet: The primary antibodies against β-actin (81115-1-RR), APC (19782-1-AP), Dvl1 (27384-1-AP), β-catenin (51067-2-AP), HRP-Goat Anti-Rabbit IgG (SA00001-2) were purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques:

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 2 | DVL1 knockdown reduces β-catenin levels in the cartilage and chondrocytes. Mice with OA were administered adenoviral vectors containing either sh-DVL1 or sh-NC via intra-articular injections. Lentiviral vectors carrying sh-DVL1 or sh-NC were introduced into chondrocytes extracted from OA mice. The mRNA (A) and protein (B) levels of DVL1 in chondrocytes were determined using RT-qPCR and WB analysis, respec- tively. (C) Protein levels of β-catenin in chondrocytes determined using WB analysis. (D) The transcriptional activity of β-catenin in chondrocytes analyzed by TOP/FOPFlash assays. (E) Positive staining of DVL1 in the mouse knee joint determined using immunofluorescence staining. (F) mRNA expression of DVL1 in the joint cartilage determined using RT-qPCR. (G) Positive expression of β-catenin in the joint cartilage determined us- ing IHC assay. For animal experiments, each group contained five mice. For cell experiments, three biological replicates were performed. Differences were compared by the unpaired t-test (C, D) or ANOVA (A, B, F, and G).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Knockdown, Quantitative RT-PCR, Activity Assay, Staining, Immunofluorescence, Expressing

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 3 | DVL1 knockdown ameliorates cartilage injury in mice. (A) Protein levels of COL10A1, MMP13, and SOX9 in mouse cartilage tissues determined using WB analysis. (B) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (C) Positive TRAP staining in the mouse knee joints. (D) Apoptosis in the extracted chondrocytes determined using TUNEL assay. (E) Protein levels of pro- cleaved-caspase-3 in the extracted chondrocytes determined using WB analysis. (F) Protein levels of COL10A1, MMP13, and SOX9 in the mouse chondrocytes determined using WB analysis. For animal experiments, each group contained five mice. For cell experiments, three biological repli- cates were performed. Differences were compared by the unpaired t-test (D–F) or ANOVA (A–C).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Knockdown, Staining, TUNEL Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 4 | CHIR-99021 restores cartilage injury mitigated by DVL1 silencing. Mice stably administered sh-DVL1 were further treated with the Wnt/β-catenin agonist CHIR-99021 via intra-articular injection. (A) Protein levels of β-catenin in cells determined using WB analysis. (B) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (C) Protein levels of COL10A1 and COL2A1 in mouse chon- drocytes determined using WB analysis. (D) Positive TRAP staining in the mouse knee joints; in vitro, chondrocytes with stable DVL1 knockdown were treated with 5 μM CHIR-99021 for 24 h. (E) Apoptosis in the extracted chondrocytes determined using TUNEL assay. (F) Protein levels of pro- cleaved-caspase-3 in the extracted chondrocytes determined using WB analysis. (G) Protein levels of expression of MMP13 and SOX9 in the mouse chondrocytes determined using WB analysis. For animal experiments, each group contained five mice. For cell experiments, three biological repli- cates were performed. Differences were compared by the unpaired t-test (A–G).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Stable Transfection, Injection, Staining, In Vitro, Knockdown, TUNEL Assay, Expressing

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 5 | SP2, poorly expressed in OA, represses DVL1 transcription. (A) Transcription factor binding sites near the mouse DVL1 promoter region predicted using the JASPAR Transcription Factors plugin from the UCSC Genome Browser. (B) Intersections of the predicted transcription factors and the significant DEGs obtained from high-throughput sequencing. (C) Positive staining of SP2 in the joint cartilage determined using IHC. (D) SP2 mRNA expression in the joint cartilage determined using RT-qPCR. (E) Binding between SP2 and the DVL1 promoter in chondrocytes determined using ChIP-qPCR assay; the extracted chondrocytes were administered lentiviral vectors carrying OE-NC or OE-SP2. (F) mRNA and (G) protein level of SP2 in mouse chondrocytes determined using RT-qPCR and WB analysis. (H) mRNA expression of DVL1 in mouse chondrocytes analyzed using RT-qPCR. (I) Regulation of SP2 on transcription activity of the DVL1 promoter in chondrocytes determined using the dual lucifer- ase reporter gene assay. For animal experiments, each group contained five mice. For cell experiments, three biological replicates were performed. Differences were compared by the unpaired t-test (C–I).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Binding Assay, Next-Generation Sequencing, Staining, Expressing, Quantitative RT-PCR, ChIP-qPCR, Activity Assay, Reporter Gene Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 6 | DVL1 upregulation aggravates cartilage injury in mice alleviated by SP2. DMM-challenged mice were introduced with adenoviral vectors carrying OE-NC/OE-SP2 alone or with the additional OE-NC/OE-DVL1. (A) mRNA expression of SP2 and DVL1 in the joint cartilage deter- mined using RT-qPCR. (B) Positive staining of SP2 and DVL1 in the joint cartilage determined using IHC. (C) Protein levels of β-catenin in the joint cartilage determined using WB analysis. (D) Cartilage morphology in the mouse knee joints determined using Safranin O/fast green staining. (E) Positive TRAP staining of the mouse knee joints. (F) Protein levels of COL10A1 and MMP13 in the mouse cartilage determined using WB analysis. Each group contained five mice. Differences were compared by ANOVA (A–F).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Expressing, Quantitative RT-PCR, Staining

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 7 | DVL1 overexpression increases chondrocyte loss suppressed by SP2. Chondrocytes were administered lentiviral vectors encapsu- lating OE-NC/OE-SP2 or the additional OE-NC/OE-DVL1. (A) mRNA expression of DVL1 in cells determined using RT-qPCR. (B) Apoptosis in cells determined using TUNEL assay. (C) Protein levels of β-catenin in chondrocytes determined using WB analysis. (D) Transcriptional activity of β-catenin in chondrocytes analyzed by TOP/FOPFlash assays. (E) Protein levels of SOX9 and COL2A1 in the chondrocytes determined using WB analysis. Three biological replicates were performed. Differences were compared by unpaired t-test (A) or ANOVA (B–E).

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, TUNEL Assay, Activity Assay

Journal: The journal of gene medicine

Article Title: Sp2 Transcription Factor Alleviates Chondrocyte Loss in Osteoarthritis by Repressing the DVL1-Dependent Wnt/β-Catenin Signaling Pathway.

doi: 10.1002/jgm.70021

Figure Lengend Snippet: FIGURE 8 | Schematic illustration of the mechanism. In cartilage tissues of osteoarthritic mice, decreased expression of SP2 resulted in its attenuated transcriptional repression of DVL1. Aberrantly expressed DVL1 activated the β-catenin signaling, leading to chondrocyte damage and osteoarthritis progression.

Article Snippet: The sections were blocked with 5% goat serum for 1 h, followed by an overnight incubation at 4°C with the antibodies against DVL1 (1:100, 13–706, ProSci, Poway, California, United States), β- catenin (1:500, ab32572, Abcam), and SP2 (1:100, PA5- 103254, Thermo Fisher Scientific).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: miR-23a promotes IKK α expression but suppresses ST7L expression to contribute to the malignancy of epithelial ovarian cancer cells

doi: 10.1038/bjc.2016.244

Figure Lengend Snippet: miR-23a positively regulates the NF- κ B and WNT/MAPK pathways. ( A – D ) OVCAR3 cells were transfected with the indicated combinations of pri-miR-23a, ASO-miR-23a, pcDNA3/IKK α , pri-miR-23a, and shR-IKK α . After 48 h, cells were harvested, and p50 was detected by immunofluorescent staining and western blotting. n-p50: nuclear p50, the control group was CENPA; c-p50: cytoplasmic p50, the control group was GAPDH (glyceraldehyde 3-phosphate dehydrogenase). ( E ) Western blotting were used to measure the protein levels after transfection with pri-miR-23a or ASO-miR-23a. ( F ) Western blotting showed the protein levels in cells treated with pcDNA3/ST7L or shR-ST7L. ( G ) OVCAR3 and SKOV3 cells were cotransfected with pTOP/flash or pFOP/flash, pcDNA3, pri-miR-23a, pcDNA3/ST7L, or pri-miR-23a and pcDNA3/ST7L, respectively, after which luciferase reporter assays were performed. ( H ) The model by which miR-23a upregulates IKK α and downregulates ST7L to activate NF- κ B and WNT/MAPK pathways to promote tumour malignancy. All of the experiments were repeated three times. * P <0.05; ** P <0.01; *** P <0.001; NS, not significant.

Article Snippet: Antibodies of ICAM-1, Vimentin, E-cadherin, wnt2, Dvl1, MAP3K7, β -catenin, GAPDH, CENPA, ST7L, and IKK α were purchased from Tianjin Saier Biotech (Tianjin, China) and the secondary goat anti-rabbit antibody was obtained from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Transfection, Staining, Western Blot, Control, Luciferase

Journal: Oncology Reports

Article Title: Expression of Wnt pathway components frizzled and disheveled in colon cancer arising in patients with inflammatory bowel disease

doi: 10.3892/or.18.3.691

Figure Lengend Snippet: Figure 2. DVL2 (A-C) and DVL3 (D-F) expression by immunohistochemistry in representative tissue samples from patients with inflammatory bowel disease and colon cancer. Expression of DVL2 and DVL3 is seen in the normal (IBD) colonic mucosa (A and D, arrows) and dysplastic areas adjacent to the colon cancer (B and E, arrows). No expression is seen in colon cancer tissue (C and F).

Article Snippet: Primary antibodies included polyclonal goat antihuman Fz1+2 antibodies, which react with both human Fz1 and Fz2 cell surface receptors (clone sc-7429, Santa Cruz Biotechnology, Santa Cruz, CA) and antibodies specific for DVL1 (clone Ab5970, rabbit polyclonal, Chemicon Corp, Temecula, CA), DVL2 (clone sc-7399, goat polyclonal, Santa Cruz Biotechnology) and DVL 3 (clone sc-26504, goat polyclonal, Santa Cruz Biotechnology).

Techniques: Expressing, Immunohistochemistry