Journal: Cell Reports

Article Title: Evolutionary loss of inflammasomes in the Carnivora and implications for the carriage of zoonotic infections

doi: 10.1016/j.celrep.2021.109614

Figure Lengend Snippet:

Article Snippet: MiSeq Reagent Kit v2 , Illumina , Cat#MS-103-1001.

Techniques: Virus, Recombinant, Transfection, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Reverse Transcription, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Cytotoxicity Assay, Sequencing, Real-time Polymerase Chain Reaction, Software, CRISPR

Journal: Investigative Ophthalmology & Visual Science

Article Title: Regulation of Cdc42 Expression and Signaling Is Critical for Promoting Corneal Epithelial Wound Healing

doi: 10.1167/iovs.13-11955

Figure Lengend Snippet: Effect of silencing of Cdc42 expression on corneal epithelial wound healing. RNA interference by siRNA transfection was performed in RCEC as described in Methods. The cells were treated with transfection reagent (Trfc) mixed with Cdc42 siRNA or Trfc alone for 16 to 20 hours. The cultures were incubated for an additional 24 hours in DMEM/F12 containing 1% FCS. After that, the cultures were harvested for determining Cdc42 expression levels by Western immunoblotting or subjected to scratch injury that caused the debridement of epithelium. Wounded cultures were further incubated in DMEM/F12 containing 1% FCS for 24 hours for healing to occur. (A) The images of wounded cultures captured at 0 hour and after 24 hours (same regions). The white dashed lines indicate the leading edge of the wound at 0 hour. (B) The quantification of wounded areas was performed by image analysis (Zeiss-Axiovision 4.5 software). Histograms for each experimental condition represent the covered wound area after 24 hours (measured from the 0-hour leading edge). The permanent, deep, canyon-like markings appearing on the culture dishes (right three panels in (A) did not cause impediment of cell growth. The details of image acquisition and quantification of wound healing are described in Methods. Cdc42 protein levels (C) in cultures were quantified by densitometry (D). Data shown in (B) and (D) are mean ± SD of three separate experiments (*P < 0.05 control versus various treatments as indicated).

Article Snippet: Protein A agarose beads, Cdc42 siRNA and siRNA transfection kit were from Santa Cruz Biotechnology.

Techniques: Expressing, Transfection, Incubation, Western Blot, Software, Control