Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Tianma Gouteng Decoction regulates oxidative stress and inflammation in AngII-induced hypertensive mice via transcription factor EB to exert anti-hypertension effect.

doi: 10.1016/j.biopha.2021.112383

Figure Lengend Snippet: Fig. 4. The effect of TGD on oxidative stress, endothelial inflammation, and TFEB expression in AngII-induced hypertensive mice. (A-D) Serum levels of NO, SOD, MDA, and GSH-PX after six weeks of TGD treatment. (E-F) The mRNA levels of HO1 and SOD2. (G-L) The mRNA expression levels of TNF-α, IL-1β, IL-6, SELE, VCAM- 1, and MCP-1. (M) The expression of CD31 and VCAM-1 in the aorta using the immunofluorescence method. (N) The protein expression of TFEB in the aorta was detected by immunofluorescence technology. (O) The mRNA expression of TFEB. Data are shown as mean ± SD, n = 5. *P < 0.05 vs Sham+saline; * *P < 0.01 vs Sham+saline; #P < 0.05 vs AngII+saline; ##P < 0.01 vs AngII+saline.

Article Snippet: The primary antibodies of CD31, TFEB, and the VCAM-1and secondary antibodies were purchased from Proteintech Inc. (IL, USA).

Techniques: Expressing, Immunofluorescence, Saline

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Tianma Gouteng Decoction regulates oxidative stress and inflammation in AngII-induced hypertensive mice via transcription factor EB to exert anti-hypertension effect.

doi: 10.1016/j.biopha.2021.112383

Figure Lengend Snippet: Fig. 8. After TFEB knockdown, the effects of TGD on endothelial inflammation and TFEB-PGC-1α-PPARγ signaling pathway in AngII-induced hypertensive mice. (A) The heat map of TNF-α, IL-1β, IL-6, SELE, VCAM-1, and mRNA expression: the darker the color, the lower the Cq value; the brighter the color, the higher the Cq value; (B) The mRNA expression of TFEB, PGC-α, PPARγ, and eNOS; (C) Representative images of co-stained immunofluorescence with TFEB and VCAM-1. Data are shown as mean ± SD, n = 3. *P < 0.05 vs Sham+saline; * *P < 0.01 vs Sham+saline; #P < 0.05 vs AngII+saline; ##P < 0.01 vs AngII+saline.

Article Snippet: The primary antibodies of CD31, TFEB, and the VCAM-1and secondary antibodies were purchased from Proteintech Inc. (IL, USA).

Techniques: Knockdown, Expressing, Staining, Immunofluorescence, Saline

Journal: Bioorganic & Medicinal Chemistry

Article Title: Synthesis and characterization of tritylthioethanamine derivatives with potent KSP inhibitory activity

doi: 10.1016/j.bmc.2011.07.054

Figure Lengend Snippet: Figure 4. Dose-dependent depletion of KSP from monopolar spindles in 1 and 5-treated cells. HeLa cells were incubated in either 1 (A–I) or 5 (J–R) for 4 h and the fixed and processed for KSP (green), the microtubule-binding protein EB1 (red) and DNA (blue) localization, and images were acquired using equivalent exposure times for the green channel.

Article Snippet: Cells were then probed with either mouse anti-tubulin (Sigma, Co, St. Louis, MO), or mouse anti-KSP (Abcam, Cambridge, MA) in blocking buffer overnight at 4 C. To counterstain the microtubule cytoskeleton in KSP localization experiments, cells were also probed for the presence of the microtubule end-binding protein EB1 using a custom rabbit polyclonal antibody against whole recombinant human EB1 (Pro-Sci, Poway, CA).

Techniques: Incubation, Binding Assay

Journal: bioRxiv

Article Title: A hierarchical GBP network promotes cytosolic LPS recognition and sepsis

doi: 10.1101/2021.08.25.457662

Figure Lengend Snippet: (A) . Potential positions of Gbp2 and Gbp3 in the non-canonical inflammasome pathway. (B) . Secreted Gsdmd-N, IL-1β p17 and caspase-1 p20 after iLPS exposure in BMDMs. Sup, supernatants. WCL, whole cell lysates. Representative of 3 independent repeats. (C) . Gsdmd-N, IL-1β p17 and caspase-1 p20 in genetically complemented Gbp3 - / - BMDMs. Representative of 2 independent repeats. (D) . Gsdmd-N in BMDM membrane extracts after iLPS exposure. Na + /K + ATPase, membrane marker; Gapdh, soluble marker. Representative of 4 independent repeats. (E) . Neutravidin pull-down (PD) of biotinylated PM immunoblotted for Gsdmd-N. Input, total cell lysates. Representative of 4 independent repeats. (F) . Gsdmd-N in biotinylated PM fractions 2h after iLPS transfection of complemented BMDMs. Input, total cell lysates. Representative of 2 independent repeats. (G) . PM-captured Gsdmd-N 2h after iLPS transfection in BMDMs from all 7 Gbp genotypes plus WT and Casp11 - / - controls. Representative of 2 independent repeats. (H) . SIM imaging of doxycycline-expressed Gsdmd-N and Gbp3 in HeLa cells with PM biotinylation. Insets shown on right. (I) . Cryo-immunoelectron microscopy (cryo-IEM) of Gsdmd-N and Gbp3. Sections labelled with anti-RFP and anti-GFP antibodies conjugated to 5nm and 25nm gold (Au) particles, respectively. Inset, right.

Article Snippet: LPS-primed BMDMs (5×10 5 cells) in 60-mm dish were transfected with 4μg of biotinylated-LPS (LPS EB-Biotin, Invivogen).

Techniques: Membrane, Marker, Transfection, Imaging, Immuno-Electron Microscopy

Journal: bioRxiv

Article Title: A hierarchical GBP network promotes cytosolic LPS recognition and sepsis

doi: 10.1101/2021.08.25.457662

Figure Lengend Snippet: (A). Four-color SIM imaging of RFP-Gbp3 and Gsdmd-N-GFP in transfected HeLa cells. Plasma membrane labeling used biotin-Cy5 and shown in z,y planar view (below). DAPI was used to delineate the nucleus. (B). Individual SIM channels for Gsdmd-N, Gbp3 and biotinylated plasma membrane along with overlay containing the DAPI-labelled nucleus from the insets in a,b. Dotted lines depict z-plane boundaries for the cell along with plasma membrane edge. (C). Violin plots of Gbp2 or Gbp3 colocalization with Gsdmd-N identifed via unbaised computational sampling. One-way ANOVA with Dunnett’s multiple comparison test between the two Gbp-labelled groups.

Article Snippet: LPS-primed BMDMs (5×10 5 cells) in 60-mm dish were transfected with 4μg of biotinylated-LPS (LPS EB-Biotin, Invivogen).

Techniques: Imaging, Transfection, Membrane, Labeling, Sampling, Comparison