bsm 33281m bioss rabbit anti beta catenin polyclonal antibody  (Bioss)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Brucella melitensis clinical isolate modulates osteoclast differentiation to drive pathological bone destruction in brucellar arthritis

doi: 10.3389/fcimb.2025.1694633

Figure Lengend Snippet: B. melitensis biovar 3 invades all types of bone tissue cells with MOI-dependent effects on osteocyte viability and exhibits stronger tropism for osteoclasts compared to osteoblasts and osteocytes. (A) Intracellular bacterial loads (CFU) in primary monocytes infected with clinical isolates and reference virulent and attenuated strains at different MOIs and time points. Data represent mean ± SD. One-way ANOVA with Tukey’s post-hoc test; * P < 0.05, ** P < 0.01. (B) Viability of different bone tissue cells under various MOI infection conditions. Data represent mean ± SD. One-way ANOVA with Tukey’s post-hoc test; * P < 0.05, ** P < 0.01. (C) Intracellular CFU counts in different bone tissue cells at MOI = 400 across time points. Data represent mean ± SD. One-way ANOVA with Tukey’s post-hoc test; ** P < 0.01. (D) Representative immunofluorescence micrographs of various cells 72 h post-infection at MOI = 400. Scale bar: 10μm. Blue: nuclei; red: Brucella ; green: β-catenin. (E) Quantitative analysis of Brucella red fluorescence intensity. Data represent mean ± SD. One-way ANOVA with Tukey’s post-hoc test; ** P < 0.01.

Article Snippet: Cells were blocked with Immunol Staining Blocking Buffer (Beyotime, Shanghai, China) and then incubated overnight at 4 °C with in-house goat anti- Brucella polyclonal and rabbit anti- β -catenin monoclonal (Proteintech, Wuhan, China) primary antibodies diluted 1:200 in primary antibody dilution buffer (Beyotime, Shanghai, China).

Techniques: Infection, Immunofluorescence, Fluorescence

Journal: eLife

Article Title: Wnt induces FZD5/8 endocytosis and degradation and the involvement of RSPO-ZNRF3/RNF43 and DVL

doi: 10.7554/eLife.103996

Figure Lengend Snippet: ( A ) Flow cytometry analysis of FZD levels on the cell surface of WT HEK293A, ZRDKO (ZRDKO C1/2), and HEK293A cells stably expressing V5-FZD5/7 with an anti-pan-FZD monoclonal antibody. FZD levels on the cell surface were greater in ZRDKO cells than in WT cells but much lower than those in V5-FZD5/7-expressing cells. ( B ) Cytosolic β-catenin levels were significantly lower in V5-FZD5/7-expressing cells than in ZRDKO cells. ( C ) ZNRF3/RNF43 double knockout-induced DVL phosphorylation and cytosolic β-catenin accumulation depend on endogenous Wnt proteins. WT or ZRDKO cells were treated with or without IWP-2 and analyzed by immunoblotting. ( D ) Overexpressing V5-FZD5 or V5-FZD7 resulted in Wnt-independent DVL phosphorylation. V5-FZD5/7-expressing cells were treated with or without IWP-2, and the WCLs were analyzed by immunoblotting. ( E ) Expression of ZNRF3 or RNF43, but not RNF130 or RNF150, reversed the increase in cytosolic β-catenin levels in ZRDKO cells. ( F ) Overexpression of RNF43 or RNF150 reduced the levels of mature forms of V5-FZD5 or V5-FZD7. WCLs from HEK293A cells expressing V5-FZD5 or V5-FZD7 alone or together with RNF43-HA or RNF150-HA were analyzed by immunoblotting. ( G ) Overexpression of ZNRF3, RNF43, RNF130, or RNF150 in ZRDKO cells reduced FZD levels on the cell surface, as measured by flow cytometry with an anti-pan-FZD antibody. ( H ) Overexpression of ZNRF3, RNF43, RNF130, or RNF150 in ZRDKO cells reduced FZD5/8 levels on the cell surface. IWP-2 partially reversed the reduction in FZD5/8 levels induced by ZNRF3 or RNF43 but not by RNF130 or RNF150. The cells in ( E ) were treated with or without IWP-2 overnight and analyzed by flow cytometry with an anti-FZD5/8 monoclonal antibody. Figure 5—source data 1. Raw unedited blots for . Figure 5—source data 2. Uncropped and labeled blots for .

Article Snippet: Antibody , Rabbit polyclonal anti-Beta Catenin , Proteintech , Cat# 51067–2-AP; RRID: AB_2086128 , WB(1:10000).

Techniques: Flow Cytometry, Stable Transfection, Expressing, Double Knockout, Phospho-proteomics, Western Blot, Over Expression, Labeling

Journal: eLife

Article Title: Wnt induces FZD5/8 endocytosis and degradation and the involvement of RSPO-ZNRF3/RNF43 and DVL

doi: 10.7554/eLife.103996

Figure Lengend Snippet: ( A ) Flow cytometry analysis of FZD5/8 levels on the cell surface in ZRDKO and ZRDKO-FZD5/8DKO cells with an anti-FZD5/8 antibody. ( B ) Depletion of FZD5/8 diminished the increase in cytosolic β-catenin levels in ZRDKO cells. Cytosolic fractions from the indicated cells were analyzed by immunoblotting. ( C ) Re-expression of V5-FZD5, but not V5-FZD7, restored cytosolic β-catenin levels in ZRDKO-FZD5/8DKO cells. ( D ) Overexpression of V5-FZD, but not V5-FZD7, further elevated cytosolic β-catenin levels in ZRDKO cells. ( E, F ) FZD5, but not FZD7, enhanced the inhibitory effect of RNF43 on Wnt3a signaling. HEK293A cells stably expressing RNF43-HA alone or with V5-FZD5 or V5-FZD7 were treated with increasing doses of Wnt3a for 2 hr, and the cytosolic fractions were analyzed by immunoblotting. ( G ) Flow cytometry analysis of cell surface FZD5/8 levels in WT or FZD5/8 DKO cells. ( H, I ) FZD5/8 double knockout abolished RSPO1-induced cytosolic β-catenin accumulation but had little effect on Wnt3a-induced increases in cytosolic β-catenin levels. The cells were treated with RSPO1 CM for 4 hr ( H ) or increasing doses of Wnt3a for 2 hr ( I ), and the cytosolic fractions were analyzed by immunoblotting. ( J ) WCLs from WT, FZD5/8 DKO and FZD5/8 DKO cells stably expressing V5-FZD5 or V5-FZD8 were analyzed by immunoblotting. ( K ) FZD5/8 double knockout abolished RSPO1-induced cytosolic β-catenin accumulation, which was restored by re-expressing V5-FZD5 or V5-FZD8. Figure 6—source data 1. Raw unedited blots for . Figure 6—source data 2. Uncropped and labeled blots for .

Article Snippet: Antibody , Rabbit polyclonal anti-Beta Catenin , Proteintech , Cat# 51067–2-AP; RRID: AB_2086128 , WB(1:10000).

Techniques: Flow Cytometry, Western Blot, Expressing, Over Expression, Stable Transfection, Double Knockout, Labeling