Journal: iScience

Article Title: A small-molecule Skp1 inhibitor elicits cell death by p53-dependent mechanism

doi: 10.1016/j.isci.2022.104591

Figure Lengend Snippet:

Article Snippet: Β-actin-HRP Rabbit Monoclonal antibody , Beyotime , Cat# AF5006.

Techniques: Control, Virus, Recombinant, Synthesized, Protease Inhibitor, Transfection, Marker, Membrane, Caspase-Glo Assay, Protein Extraction, Bicinchoninic Acid Protein Assay, Western Blot, Plasmid Preparation, SYBR Green Assay, Software

Journal: Cell Communication and Signaling : CCS

Article Title: LRRK2 is involved in the chemotaxis of neutrophils and differentiated HL-60 cells, and the inhibition of LRRK2 kinase activity increases f MLP-induced chemotactic activity

doi: 10.1186/s12964-023-01305-y

Figure Lengend Snippet: Lrrk2 knockdown results in a decrease in OXPHOS activities of dHL-60 cells. A , B dHL-60 cells expressing tetracycline-inducible Lrrk2 shRNA or irrelevant (Irr) shRNA were cultured in 1.25% DMSO with 0.5 µg/ml doxycycline (Dox) for 3 days. The OXPHOS activities of these cells were measured after treatment with 1 µM f MLP for 5 min ( n = 4). Error bars, SEM. A complex I. B complex I + II. C Representative immunofluorescence microscope images of mitochondria. dHL-60 cells expressing tetracycline-inducible Lrrk2 shRNA or irrelevant (Irr) shRNA were cultured in 1.25% DMSO with 0.5 µg/ml Dox for 3 days. dHL-60 cells attached to coverslips were then incubated with or without 10 nM f MLP for 5 min. Green; anti-TOMM20, Red; anti-ß-actin, Blue; DAPI. Bar, 10 µm. D Percentages of cells with mitochondria of tubular form after f MLP stimulation. > 50 cells were analyzed in four independent experiments. Error bars, SEM. ** p < 0.01 compared with Irr shRNA expressing dHL-60 cells

Article Snippet: The following antibodies were used in this study: mouse monoclonal antibodies against ß-actin (Merck) and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit monoclonal antibodies against MFN2 (Abcam, Tokyo, Japan), LRRK2 (Abcam), LRRK2 pS935 (Abcam), RAB10 (Abcam), RAB10 pT73 (Abcam), and TOMM20 (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, shRNA, Cell Culture, Immunofluorescence, Microscopy, Incubation

Journal: PLoS ONE

Article Title: A Novel Zebrafish Model to Provide Mechanistic Insights into the Inflammatory Events in Carrageenan-Induced Abdominal Edema

doi: 10.1371/journal.pone.0104414

Figure Lengend Snippet: (A) Western blots for MPO, TNF-α, iNOS, and β-actin proteins from adult zebrafish; (B) relative density of the MPO immunoblot; (C) relative density of the TNF-α immunoblot; (D) relative density of the iNOS immunoblot. There are 3 groups: vehicle+vehicle: i.p. injection first vehicle (20 µL PBS) 1 h before i.p. second vehicle (20 µL PBS); vehicle+carra: i.p. vehicle (20 µL PBS) 1 h before i.p. 1.5% carrageenan (20 µL); MP+carra: i.p. 1 µg MP (20 µL) 1 h before i.p. 1.5% carrageenan (20 µL). The abdominal samples from adult zebrafish were collected at 24 h after the second injection (as well as 25 h after the first injection). Relative intensity of the i.p. vehicle plus i.p. 1.5% carrageenan group was taken to be 100%. Band intensities of MPO, TNF-α, and iNOS were quantified by densitometry and are indicated as a percentage change relative to that of the vehicle plus carrageenan group. Western blotting of the abdominal tissue homogenate revealed that injection of carrageenan evoked significant upregulation of MPO, TNF-α, and iNOS proteins at 24 h compared with that of the vehicle plus vehicle group. MP administered at a dose of 1 µg (i.p.) 1 h prior to i.p. 1.5% carrageenan significantly reduced carrageenan-induced upregulation of MPO, TNF-α, and iNOS. Western blotting of β-actin was performed to verify that equivalent amounts of protein were loaded in each lane. These experiments were repeated 3 times. Ca: carrageenan; MP: methylprednisolone; MPO: myeloperoxidase, TNF-α: tumor necrosis factor-α; iNOS: inducible nitric oxide synthase. * P <0.05 compared with the i.p. vehicle plus i.p. vehicle group; # P <0.05 compared with the i.p. vehicle plus i.p. 1.5% carrageenan group.

Article Snippet: At the end, the PVDF membranes were reprobed with an anti-β-actin antibody (1∶2500 dilution; GeneTex, San Antonio, TX, USA; catalog no. GTX124500; polyclonal rabbit antibody).

Techniques: Western Blot, Injection