anti active β catenin 8814s  (Cell Signaling Technology Inc)

Journal: Journal of inherited metabolic disease

Article Title: Liver-Directed Gene Therapy Mitigates Early Nephropathy in Murine Glycogen Storage Disease Type Ia.

doi: 10.1002/jimd.70048

Figure Lengend Snippet: FIGURE 3 | Assessment of kidney AKI and fibrosis in L-G6PC1-low and L-G6PC1-high mice. L-G6PC1-low and L-G6PC1-high mice were gen- erated from G6pc−/− mice and analyzed at 12 weeks of age. Age-matched G6pc+/+ and G6pc+/− mice with a similar phenotype served as controls. (A) Western-blot analyzes and quantitation of renal levels of E-cadherin and N-cadherin in control (n = 20), L-G6PC1-low (n = 23), and L-G6PC1-high (n = 23) mice. (B) Western-blot analyzes and quantitation of renal levels of Dkk3 and CTGF in control (n = 20), L-G6PC1-low (n = 23), and L-G6PC1- high (n = 23) mice. (C) Western-blot analyzes and quantitation of renal levels of total (β-catenin-T) and active, dephosphorylated (β-catenin-A) β-catenin in control (n = 20), L-G6PC1-low (n = 23), and L-G6PC1-high (n = 23) mice. For Western-blot analyzes, densitometric quantification was performed and normalized against β-Actin. Values represent the mean ± SEM. *p < 0.05, **p < 0.005. (D) Immunohistochemical analysis of renal lev- els of active β-catenin. Scale bar, 50 μm. (E) Quantitation of renal levels of nuclear localized active β-catenin in control (n = 3), L-G6PC1-low (n = 3), and L-G6PC1-high (n = 3) mice. Kidney sections were immunostained with HRP-labeled anti-active β-catenin and the nuclei counterstained with hematoxylin. Images were digitized using the Motic EasyScan Infinity 60 scanner and analyzed with QuPath software (v0.4.3). Multiple annotations were selected across the entire renal cortex. The Nucleus DAB OD mean scoring method [19] was used to identify moderate and strong optical density thresholds for nuclear-stained β-catenin.

Article Snippet: Nuclear- translocated active β- catenin was analyzed by immunohistochemistry on paraffin- embedded mouse kidney sections using a rabbit monoclonal antibody specific for nonphosphorylated (active) β- catenin (#8814, Cell Signaling Technology).

Techniques: Western Blot, Quantitation Assay, Control, Immunohistochemical staining, Labeling, Software, Staining

Journal: Journal of Inherited Metabolic Disease

Article Title: Liver‐Directed Gene Therapy Mitigates Early Nephropathy in Murine Glycogen Storage Disease Type Ia

doi: 10.1002/jimd.70048

Figure Lengend Snippet: Assessment of kidney AKI and fibrosis in L‐G6PC1‐low and L‐G6PC1‐high mice. L‐G6PC1‐low and L‐G6PC1‐high mice were generated from G6pc −/− mice and analyzed at 12 weeks of age. Age‐matched G6pc +/+ and G6pc +/− mice with a similar phenotype served as controls. (A) Western‐blot analyzes and quantitation of renal levels of E‐cadherin and N‐cadherin in control ( n = 20), L‐G6PC1‐low ( n = 23), and L‐G6PC1‐high ( n = 23) mice. (B) Western‐blot analyzes and quantitation of renal levels of Dkk3 and CTGF in control ( n = 20), L‐G6PC1‐low ( n = 23), and L‐G6PC1‐high ( n = 23) mice. (C) Western‐blot analyzes and quantitation of renal levels of total (β‐catenin‐T) and active, dephosphorylated (β‐catenin‐A) β‐catenin in control ( n = 20), L‐G6PC1‐low ( n = 23), and L‐G6PC1‐high ( n = 23) mice. For Western‐blot analyzes, densitometric quantification was performed and normalized against β‐Actin. Values represent the mean ± SEM. * p < 0.05, ** p < 0.005. (D) Immunohistochemical analysis of renal levels of active β‐catenin. Scale bar, 50 μm. (E) Quantitation of renal levels of nuclear localized active β‐catenin in control ( n = 3), L‐G6PC1‐low ( n = 3), and L‐G6PC1‐high ( n = 3) mice. Kidney sections were immunostained with HRP‐labeled anti‐active β‐catenin and the nuclei counterstained with hematoxylin. Images were digitized using the Motic EasyScan Infinity 60 scanner and analyzed with QuPath software (v0.4.3). Multiple annotations were selected across the entire renal cortex. The Nucleus DAB OD mean scoring method was used to identify moderate and strong optical density thresholds for nuclear‐stained β‐catenin.

Article Snippet: Nuclear‐translocated active β‐catenin was analyzed by immunohistochemistry on paraffin‐embedded mouse kidney sections using a rabbit monoclonal antibody specific for non‐phosphorylated (active) β‐catenin (#8814, Cell Signaling Technology).

Techniques: Generated, Western Blot, Quantitation Assay, Control, Immunohistochemical staining, Labeling, Software, Staining

Journal: BMC oral health

Article Title: Lipid droplet accumulation impairs osseointegration by disturbing the osteogenesis-osteoclasis balance on titanium implant surface in hyperlipidemia.

doi: 10.1186/s12903-025-06218-5

Figure Lengend Snippet: Fig. 1 Evaluation of the levels of oxidative damage and Wnt/β-catenin pathway around the implants in hyperlipidemia mice. (A) Illustration of the first part of animal experimental design. (B-G) Various physiological and biochemical indicators in NC and HF group. (H) Fluorescent images of the nucleus (blue), 8-OHdG (green), and non-p-β-catenin (red) around the titanium implants in NC and HF group. (I) Quantitative analysis of mean fluorescence in tensity of 8-OHdG and non-p-β-catenin. Data was presented as mean ± SD, n = 3 specimens/group, *P < 0.05. NC: negative control; HF: high-fat; TC, total cholesterol; TG, triglycerides; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; GLU, glucose; IM: implant; DAPI, 4’, 6-diamidino-2-phenylindole; 8-OHdG, 8-hydroxy-2 deoxyguanosine; non-p-β-catenin: non-phospho-β-catenin

Article Snippet: Primary antibodies against 8-hydroxy-2 deoxyguanosine (8-OHdG, Santa Cruz Biotechnology, sc-393871, 1:300, USA), non-phospho-β-catenin (non-p-β-catenin, CST, #8814, 1:800, USA), alkaline phosphatase (ALP, HUABIO, ET1601-21, 1:100, China), receptor activator of nuclear factor-κ B ligand (RANKL, Proteintech, 23408-1- AP, 1:300, China), receptor activator of nuclear factor-κ B ligand (RANKL, Santa Cruz Biotechnology, sc-59982, 1:300, USA), nuclear factor-kappa B (NF-κB, Proteintech, 10745-1-AP, 1:300, China), inhibitor of NF-κB (I-κB, Proteintech, 10268-1-AP, 1:300, China), CD11b (Proteintech, 65055-1-Ig, 1:300, China), F4/80 (Santa Cruz Biotechnology, sc-52664, 1:300, USA), Gr-1 (Santa Cruz Biotechnology, sc-53515, 1:300, USA), glucose-regulated protein 78 (GRP78, Proteintech, 11587-1-AP, 1:300, China), perilipin2 (PLIN2, Proteintech, 15294-1-AP, 1:300, China) were applied.

Techniques: Fluorescence, Negative Control

Journal: BMC oral health

Article Title: Lipid droplet accumulation impairs osseointegration by disturbing the osteogenesis-osteoclasis balance on titanium implant surface in hyperlipidemia.

doi: 10.1186/s12903-025-06218-5

Figure Lengend Snippet: Fig. 3 Evaluation of titanium implant osseointegration. (A-B) 3D reconstructed images and quantitative calculation of BV/TV, TB.N, Tb.Th, and Tb.Sp by Micro-CT at 4 weeks after surgery. (C) Hard tissue slicing images of the bone-implant interface. (D) Typical images of H&E staining showing bone-implant interface in different groups. (E) Quantitative calculation of BIC%. Data was presented as mean ± SD, n = 3 specimens/group, *P < 0.05. NC: negative con trol; HF: high-fat; Wnt3a, an activator of Wnt/β-catenin pathway; NAC: N-acetyl-L-cysteine, a ROS antagonist; BV/TV, bone volume/tissue volume; TB.N, trabecular number; Tb.Th, trabecular thickness; Tb.Sp, trabecular separation; IM: implant; BIC%, percent of bone-implant contact

Article Snippet: Primary antibodies against 8-hydroxy-2 deoxyguanosine (8-OHdG, Santa Cruz Biotechnology, sc-393871, 1:300, USA), non-phospho-β-catenin (non-p-β-catenin, CST, #8814, 1:800, USA), alkaline phosphatase (ALP, HUABIO, ET1601-21, 1:100, China), receptor activator of nuclear factor-κ B ligand (RANKL, Proteintech, 23408-1- AP, 1:300, China), receptor activator of nuclear factor-κ B ligand (RANKL, Santa Cruz Biotechnology, sc-59982, 1:300, USA), nuclear factor-kappa B (NF-κB, Proteintech, 10745-1-AP, 1:300, China), inhibitor of NF-κB (I-κB, Proteintech, 10268-1-AP, 1:300, China), CD11b (Proteintech, 65055-1-Ig, 1:300, China), F4/80 (Santa Cruz Biotechnology, sc-52664, 1:300, USA), Gr-1 (Santa Cruz Biotechnology, sc-53515, 1:300, USA), glucose-regulated protein 78 (GRP78, Proteintech, 11587-1-AP, 1:300, China), perilipin2 (PLIN2, Proteintech, 15294-1-AP, 1:300, China) were applied.

Techniques: Micro-CT, Staining

Journal: BMC oral health

Article Title: Lipid droplet accumulation impairs osseointegration by disturbing the osteogenesis-osteoclasis balance on titanium implant surface in hyperlipidemia.

doi: 10.1186/s12903-025-06218-5

Figure Lengend Snippet: Fig. 4 Evaluation of the levels of oxidative damage, Wnt/β-catenin pathway, and osteogenesis around the implants after local injection of Wnt3a or NAC in hyperlipidemia mice. (A) Fluorescent images of the nucleus (blue), 8-OHdG (green) and non-p-β-catenin (red). (B) Quantitative analysis of mean fluo rescence intensity of 8-OHdG and non-p-β-catenin. (C) Fluorescent images of the nucleus (blue), 8-OHdG (green) and ALP (red). (D) Quantitative analysis of mean fluorescence intensity of 8-OHdG and ALP. Data was presented as mean ± SD, n = 3 specimens/group, *P < 0.05. NC: negative control; HF: high-fat; Wnt3a, an activator of Wnt/β-catenin pathway; NAC: N-acetyl-L-cysteine, a ROS antagonist; IM: implant; DAPI, 4’, 6-diamidino-2-phenylindole; 8-OHdG, 8-hydroxy-2 deoxyguanosine; non-p-β-catenin: non-phospho-β-catenin; ALP, alkaline phosphatase

Article Snippet: Primary antibodies against 8-hydroxy-2 deoxyguanosine (8-OHdG, Santa Cruz Biotechnology, sc-393871, 1:300, USA), non-phospho-β-catenin (non-p-β-catenin, CST, #8814, 1:800, USA), alkaline phosphatase (ALP, HUABIO, ET1601-21, 1:100, China), receptor activator of nuclear factor-κ B ligand (RANKL, Proteintech, 23408-1- AP, 1:300, China), receptor activator of nuclear factor-κ B ligand (RANKL, Santa Cruz Biotechnology, sc-59982, 1:300, USA), nuclear factor-kappa B (NF-κB, Proteintech, 10745-1-AP, 1:300, China), inhibitor of NF-κB (I-κB, Proteintech, 10268-1-AP, 1:300, China), CD11b (Proteintech, 65055-1-Ig, 1:300, China), F4/80 (Santa Cruz Biotechnology, sc-52664, 1:300, USA), Gr-1 (Santa Cruz Biotechnology, sc-53515, 1:300, USA), glucose-regulated protein 78 (GRP78, Proteintech, 11587-1-AP, 1:300, China), perilipin2 (PLIN2, Proteintech, 15294-1-AP, 1:300, China) were applied.

Techniques: Injection, Fluorescence, Negative Control

Journal: BMC oral health

Article Title: Lipid droplet accumulation impairs osseointegration by disturbing the osteogenesis-osteoclasis balance on titanium implant surface in hyperlipidemia.

doi: 10.1186/s12903-025-06218-5

Figure Lengend Snippet: Fig. 8 Oxidative damage induced by lipid droplet accumulation impairs the physiological osteogenesis-osteoclasis balance on titanium surface in hyper lipidemia. ERS and mitochondrial damage induced by lipid droplet accumulation in hyperlipidemia tends to form a vicious cycle network and promotes the expression of inflammatory factors in peri-implant tissues, which further dramatically drives osteoclastogenesis and inhibits osteogenesis on titanium implant surface. After local application of NAC, the ROS level is downregulated, which effectively activates the Wnt/β-catenin signaling pathway and in hibits the RANKL/NF-κB signaling pathway to alleviate the osteogenesis-osteoclasis imbalance, thereby ultimately facilitating the Ti implant osseointegra tion in hyperlipidemia. MDA, malondialdehyde; ROS, reactive oxygen species; ERS, endoplasmic reticulum stress; IL-1β, interleukin-1β; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; RANKL, receptor activator of nuclear factor-κ B ligand; IM, implant; NAC, N-acetyl-L-cysteine, a ROS antagonist; non-p-β- catenin, non-phospho-β-catenin; DNA, deoxyribonucleic acid; ALP, alkaline phosphatase; OPG, osteoprotegerin; COLI, collagen type I

Article Snippet: Primary antibodies against 8-hydroxy-2 deoxyguanosine (8-OHdG, Santa Cruz Biotechnology, sc-393871, 1:300, USA), non-phospho-β-catenin (non-p-β-catenin, CST, #8814, 1:800, USA), alkaline phosphatase (ALP, HUABIO, ET1601-21, 1:100, China), receptor activator of nuclear factor-κ B ligand (RANKL, Proteintech, 23408-1- AP, 1:300, China), receptor activator of nuclear factor-κ B ligand (RANKL, Santa Cruz Biotechnology, sc-59982, 1:300, USA), nuclear factor-kappa B (NF-κB, Proteintech, 10745-1-AP, 1:300, China), inhibitor of NF-κB (I-κB, Proteintech, 10268-1-AP, 1:300, China), CD11b (Proteintech, 65055-1-Ig, 1:300, China), F4/80 (Santa Cruz Biotechnology, sc-52664, 1:300, USA), Gr-1 (Santa Cruz Biotechnology, sc-53515, 1:300, USA), glucose-regulated protein 78 (GRP78, Proteintech, 11587-1-AP, 1:300, China), perilipin2 (PLIN2, Proteintech, 15294-1-AP, 1:300, China) were applied.

Techniques: Expressing