regγ antibody  (Proteintech)

Journal: Translational Gastroenterology and Hepatology

Article Title: REGγ deficiency ameliorates hepatic ischemia and reperfusion injury in a mitochondrial p66shc dependent manner in mice

doi: 10.21037/tgh-24-46

Figure Lengend Snippet: REGγ −/− inhibits hepatic mitochondrial injury in mice. In vivo experiment: Transmission electron microscopy, magnification 12,000×, scale bar 500 nm (A), Mitochondrial average diameter analysis (B); WB of p-DRP/DRP (C), gray value analysis (D); WB of Cytochrome C (E), gray value analysis (F); n=5 per group, *, P<0.05 vs. WT-Sham group; # , P<0.05 vs. WT-I/R group. In vitro experiments: JC-1 fluorescence staining 20× and MFI analysis (G,H). ROS fluorescence staining was performed 20×, and MFI analysis (I,J). n=5 per group, *, P<0.05 vs. WT-control group; # , P<0.05 vs. WT-H/R group. I/R, ischemia and reperfusion; WT, wild type; REG, the 11S proteasome regulatory complex; DRP, dynamin related protein 1; ROS, reactive oxygen species; WB, Western blot; MFI, mean fluorescence intensity; H/R, hypoxia and reoxygenation.

Article Snippet: Primary antibodies as following: 78–82 Kda hosphor-DRP1 Rabbit 1:1000, CST (Shanghai, China); 83 Kda DRP1 Rabbit 1:1000, Abcam (Shanghai, China); 61/55 Kda hosphor-NF-κB p65 Rabbit 1:1000, Bioss (Beijing, China); 65 Kda NF-κB p65 Rabbit 1:2000, Bioss; 36 Kda p-IκBα Rabbit 1:2000, CST; 36 Kda IκBα Rabbit 1:2000, CST; 35 Kda Caspase-3 Rabbit 1:1000, CST; 17/19 Kda Cleaved Caspase-3 Rabbit 1:1000, CST; 21 Kda BCL2-associated X protein (BAX) Rabbit 1:1000, Proteintech (Wuhan, China); 26 Kda B-cell lymphoma-2 (BCL-2) Rabbit 1:1000, Proteintech; 42 Kda β-actin Mouse 1:5000, Proteintech; 63/67–75 Kda p66shc Rabbit 1:1000, Proteintech; 63/55 Kda p-ser36 p66shc Mouse 1:1000, Abcam; 36 Kda GAPDH Rabbit 1:5000, Affinity (Nanjing, China); 12–15 Kda Cytochrome C Mouse 1:5000, Proteintech; 17–18 Kda COXIV Rabbit 1:5000, Proteintech; 35 REGγ Rabbit 1:2000, Proteintech.

Techniques: In Vivo, Transmission Assay, Electron Microscopy, In Vitro, Fluorescence, Staining, Control, Western Blot

Journal: mSystems

Article Title: Paneth Cells Protect against Acute Pancreatitis via Modulating Gut Microbiota Dysbiosis

doi: 10.1128/msystems.01507-21

Figure Lengend Snippet: AP patients and experimental AP mice presented Paneth cell defects. (A) Histopathological changes and mean numbers of Paneth cells per crypt of duodenal mucosa specimens were assessed by H&E staining. Original magnification, ×200 ( n = 7 to 14 individuals per group). (B) Lysozyme expression (green) was assessed in Paneth cells of duodenal mucosa specimens by immunofluorescence (magnification, ×200) and lysozyme-positive/DAPI + quantification. (C to K) The mRNA expression levels of lysozyme (C), HD6 (D), HD5 (E), Reg3γ (F), Ang4 (G), sPLA2 (H), TGFβ (I), Wnt3a (J), and Lgr5 (K) were assessed. (L) Lysozyme expression (green) (magnification, ×200) and lysozyme-positive/DAPI + quantification in three AP models. (M) Mean number of Paneth cells per crypt in AP models. The data are presented as the means ± SD. ns, no significant difference; *, P ≤ 0.05.

Article Snippet: The membrane was blocked with 3% bovine serum albumin (BSA) for 1 h and incubated with primary antibodies, diluted in primary antibody dilution buffer (Epizyme Biotech, China), against Lgr5 (catalog number A10545; Abclonal, China), lysozyme (catalog number A0099; Dako, Denmark), Reg3γ (catalog number sc-377038; Santa Cruz Biotechnology, USA), Defa5 (catalog number A18208; Abclonal, China), Ang4 (catalog number sc-377497; Santa Cruz Biotechnology, USA), and sPLA2 (catalog number sc-58363; Santa Cruz Biotechnology, USA) overnight at 4°C.

Techniques: Staining, Expressing, Immunofluorescence