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

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

Journal: eLife

Article Title: Nicotine enhances the stemness and tumorigenicity in intestinal stem cells via Hippo-YAP/TAZ and Notch signal pathway

doi: 10.7554/eLife.95267

Figure Lengend Snippet:

Article Snippet: Antibody , rabbit polyclonal anti-β-catenin , Cell Signaling Technology , #9562 , IHC (1:200).

Techniques: