Journal: Cell Reports

Article Title: SARS-CoV-2 ferritin nanoparticle vaccines elicit broad SARS coronavirus immunogenicity

doi: 10.1016/j.celrep.2021.110143

Figure Lengend Snippet:

Article Snippet: HRP-conjugated AffiniPure Goat Anti-Mouse Fcγ subclass 2c , Jackson ImmunoResearch , Cat# 115-035-208.

Techniques: Binding Assay, Virus, Recombinant, Expressing, Luciferase, Software, Plasmid Preparation, Electron Microscopy

Journal: Pharmacological research

Article Title: Apabetalone, a BET protein inhibitor, inhibits kidney damage in diabetes by preventing pyroptosis via modulating the P300/H3K27ac/PLK1 axis.

doi: 10.1016/j.phrs.2024.107306

Figure Lengend Snippet: Fig. 3. Apabetalone prevented HG-induced pyroptosis in HK-2 cells. (A) BRD4, NF-κB p-p65, NLRP3, Caspase-1 p10 and GSDMD-N protein expression. (B) FN, Collagen I protein expression. (C) The ELISA method was utilized to quantify the IL-1β levels in the supernatant. (D, E) measure the mRNA levels of IL-6 and MCP-1. (F) LDH activity. (G) cell pyroptosis by flow cytometry. (H) immunofluorescence was used to evaluate GSDMD. #P < 0.05 vs. the NG group, ##P < 0.01 vs. the NG group, *P < 0.05 vs. the HG group, **P < 0.01 vs. the HG group.

Article Snippet: Powdered apabetalone (S7295, Selleck.cn, USA) and C646 (HY-13823, MCE, USA) were dissolved in DMSO, with the final concentration 25 μM, 10 μM administered to HK-2 cells for 48 hours, respectively.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Flow Cytometry, Immunofluorescence

Journal: Pharmacological research

Article Title: Apabetalone, a BET protein inhibitor, inhibits kidney damage in diabetes by preventing pyroptosis via modulating the P300/H3K27ac/PLK1 axis.

doi: 10.1016/j.phrs.2024.107306

Figure Lengend Snippet: Fig. 4. The overall traits of db/db mice following apabetalone administration. (A) weekly fluctuations in the weights of mice. (B) blood glucose levels were evaluated during apabetalone treatment. (C, D) at 20 weeks of age old, various groups were tested for their serum creatinine and blood urea nitrogen (BUN) levels. (E) Urinary albumin to creatinine ratio (UACR) was assessed in mice when they reached 20 weeks old. (F) GBM thickness (red arrows). (G) foot process width (yellow arrows). (H) Tissue samples were performed transmission electron microscopy (TEM), stained using hematoxylin-eosin (H&E) (200×), periodic acid-Schiff (PAS)(200×) and Masson’s trichrome (Masson) staining (200×). Control: db/m mice, DKD: db/db mice. APA: db/db+Apabetalone (n=6). #P < 0.05 vs. the control group, ##P < 0.01 vs. the control group, *P < 0.05 vs. the DKD group, **P < 0.01 vs. the DKD group.

Article Snippet: Powdered apabetalone (S7295, Selleck.cn, USA) and C646 (HY-13823, MCE, USA) were dissolved in DMSO, with the final concentration 25 μM, 10 μM administered to HK-2 cells for 48 hours, respectively.

Techniques: Transmission Assay, Electron Microscopy, Staining, Control

Journal: Pharmacological research

Article Title: Apabetalone, a BET protein inhibitor, inhibits kidney damage in diabetes by preventing pyroptosis via modulating the P300/H3K27ac/PLK1 axis.

doi: 10.1016/j.phrs.2024.107306

Figure Lengend Snippet: Fig. 5. Apabetalone inhibits pyroptosis-related protein in diabetic mouse. (A) apabetalone’s chemical structure is that of a small, innovative synthetic compound within the Quinazoline group. (B) mouse experimental protocol diagram. (C, H) expression of pyroptosis-related protein and ECM proteins. (E) employed to quantify the IL-1β levels in the renal cortex homogenate of mice. (F, G) The levels of IL-6 and MCP-1 mRNA were measured. (D, I) immunohistochemical analysis of NLRP3, GSDMD, BRD4, FN and Collagen I in mouse kidney tissues (200x). Bar = 50 mm. Control: db/m mice, DKD: db/db mice. APA: db/db+Apabetalone (n=6). #P < 0.05 vs. the control group, ##P < 0.01 vs. the control group, *P < 0.05 vs. the DKD group, **P < 0.01 vs. the DKD group.

Article Snippet: Powdered apabetalone (S7295, Selleck.cn, USA) and C646 (HY-13823, MCE, USA) were dissolved in DMSO, with the final concentration 25 μM, 10 μM administered to HK-2 cells for 48 hours, respectively.

Techniques: Expressing, Immunohistochemical staining, Control

Journal: Pharmacological research

Article Title: Apabetalone, a BET protein inhibitor, inhibits kidney damage in diabetes by preventing pyroptosis via modulating the P300/H3K27ac/PLK1 axis.

doi: 10.1016/j.phrs.2024.107306

Figure Lengend Snippet: Fig. 7. In HK-2 cells, the interaction between BRD4 and PLK1 is enhanced by P300 via H3K27ac. (A) Levels of P300, H3K27ac, and PLK1 were measured following transfection with P300 siRNA. (B) After transfection with P300 siRNA, the expression levels of NF-κB p-p65, NLRP3, Caspase-1 p20, and GSDMD-N were assessed. (C) expression of extracellular matrix proteins FN and Collagen I by P300 siNRA intervention. (D, E) the silencing of P300 in HK-2 cells treated with HG led to decreased binding of H3K27ac and PLK1, as confirmed by RT-qPCR and agarose gel electrophoresis. (F) Co-IP analysis showed decreased interaction between BRD4 and H3K27ac in HK-2 cells treated with HG. (G) Levels of PLK1, H3K27ac, FN, Collagen I, NF-κB p-p65, NLRP3, Caspase-1 p10, and GSDMD-N proteins were assessed after siP300 treatment combined with enhanced BRD4 expression. (H) schematic diagram of the role of apabetalone in DKD (n=3). #P < 0.05 vs. the NG+siNC group, ##P < 0.01 vs. the NG+siNC group, *P < 0.05 vs. the HG+siNC group, **P < 0.01 vs. the HG+siNC group.

Article Snippet: Powdered apabetalone (S7295, Selleck.cn, USA) and C646 (HY-13823, MCE, USA) were dissolved in DMSO, with the final concentration 25 μM, 10 μM administered to HK-2 cells for 48 hours, respectively.

Techniques: Transfection, Expressing, Binding Assay, Quantitative RT-PCR, Agarose Gel Electrophoresis, Co-Immunoprecipitation Assay

Journal: Cerebral cortex (New York, N.Y. : 1991)

Article Title: NCAM/spectrin complex disassembly results in PSD perforation and postsynaptic endocytic zone formation.

doi: 10.1093/cercor/bhq283

Figure Lengend Snippet: Figure 9. NCAM and bI spectrin deficiency enhance AMPAR endocytosis. (A,B) Live NCAMþ/þ and NCAM/ cultured hippocampal neurons (A) or control and bI spectrin siRNA-transfected NCAMþ/þ neurons (B) were labeled with antibodies against the extracellular domain of GluR1 followed by Cy3 conjugated secondary antibodies (red). After 1 h, neurons were fixed and surface Cy3/GluR1 complexes were visualized with Cy2 conjugated secondary antibodies (green). SiRNA-transfected neurons were colabeled with bI spectrin antibodies (blue). Images of the representative neurites are shown. Note increased numbers of Cy3-positive/Cy2-negative internalized GluR1 accumulations (arrows) in NCAM/ and bI spectrin siRNA-transfected NCAMþ/þ neurons. Bar 5 10 lm. (C) Live NCAMþ/þ and NCAM/ cultured hippocampal neurons were incubated with Alexa Fluor 546-transferrin conjugates for 1 h. Representative images of neurons colabeled with MAP2 antibodies are shown. Bar 5 10 lm. Graphs (A--C) show quantitation of GluR1, GluR2, and transferrin receptor internalization (mean ± SEM, n $ 100 neurites were analyzed in each group from 45 neurons from 3 cultures). In A,C, mean internalization levels in NCAMþ/þ neurons were set to 100%. In B, mean internalization levels in control siRNA-transfected NCAMþ/þ neurons were set to 100%. *P \ 0.05, Kolmogorov--Smirnov test (for A--C).

Article Snippet: Neurons were then washed with culture media 3 times and incubated with nanogold anti-mouse-Fab conjugates (Nanoprobes) for electron microscopy or Cy3-conjugated goat anti-mouse antibodies (Jackson ImmunoResearch Laboratories, Inc.) for confocal microscopy for 15 min in a CO2 incubator.

Techniques: Cell Culture, Control, Transfection, Labeling, Incubation, Quantitation Assay