Journal: ACS Omega

Article Title: Application of an Activity-Based Probe to Determine Proteolytic Activity of Cell Surface Cathepsin G by Mass Cytometry Data Acquisition

doi: 10.1021/acsomega.0c04092

Figure Lengend Snippet: List of CyTOF Antibodies Used for the Experiments a

Article Snippet: 154 Sm , CD45 , HI30 , Fluidigm, #201302, Human PB Basic Phenotyping Kit , .

Techniques:

Journal: British Journal of Pharmacology

Article Title: The suppression of bromodomain and extra‐terminal domain inhibits vascular inflammation by blocking NF‐κB and MAPK activation

doi: 10.1111/bph.13657

Figure Lengend Snippet: Effect of BET inhibition on TNF‐α‐induced NF‐κB activation in HUVECs. (A and B) Western blot analysis of phosphorylated IKK or phosphorylated and degradated IκBα in HUVECs treated with 10 ng·mL−1 TNF‐α for 15 min in the presence of DMSO or JQ1. Data represent relative quantities obtained by densitometry (lower panel) from five independent experiments (mean ± SEM; *P < 0.05 vs DMSO, # P < 0.05 vs TNF‐α treatment; one‐way ANOVA test). (C and D) Effect of JQ1 on nuclear translocation of p65. HUVECs were treated with TNF‐α (10 ng·mL−1) for 30 min in the presence or absence of JQ1. The representative images show immunofluorescence staining analysis of p65 localization (green). Nuclei were stained with DAPI. (D ) Mean intensity of nuclear p65 protein from 5 independent experiments (mean ± SEM; *P < 0.05 vs DMSO, # P < 0.05 vs TNF‐α alone; one‐way ANOVA test). (E and F) The expression of p65 protein, detected by western blot analysis, in nuclear and cytoplasmic fractions isolated from HUVECs. Results of semiquantitative densitometry of nuclear p65 protein expression are shown in (D), from five independent experiments (mean ± SEM; *P < 0.05 vs TNF‐α alone, Student's t‐test). (G) Western blot analysis of TNF‐α‐induced phosphorylation of IKKβ in HUVECs transfected with Brd2 shRNA or Brd4 shRNA or control shRNA in the presence or absence of TNF‐α stimulation. Results of semiquantitative densitometry of protein expression are shown in the right panel, from five independent experiments (mean ± SEM; *P < 0.05 vs control shRNA, # P < 0.05 vs TNF‐α treatment; one‐way ANOVA test).

Article Snippet: Equal amounts of the samples were then separated by SDS‐PAGE gel and transferred onto NC membranes and immunoblotted with the indicated antibodies: anti‐VCAM1 (Abcam), anti‐ICAM1 (Abcam), anti‐ E‐selectin (Abcam) anti‐JNK (Cell signalling), anti‐phospho‐JNK(Cell signalling), anti‐p38(Cell signalling), anti‐phospho‐p38(Cell signalling), anti‐ERK(Cell signalling), anti‐phospho‐ERK(Cell signalling), anti‐NF‐κB p65(Cell signalling), anti‐phospho‐IκBα (Cell signalling), anti‐IκBα (Cell signalling) and anti‐phospho‐IκB kinase (IKK) (Cell signalling).

Techniques: Inhibition, Activation Assay, Western Blot, Translocation Assay, Immunofluorescence, Staining, Expressing, Isolation, Transfection, shRNA