Journal: Journal of Experimental Botany

Article Title: Unraveling the induction of phytoene synthase 2 expression by salt stress and abscisic acid in Daucus carota

doi: 10.1093/jxb/ery207

Figure Lengend Snippet: Subcellular localization of DcAREB transcription factors. Co-transformation of tobacco leaves with Agrobacterium carrying the DcAREB:RFP vectors and the 35S:GFP vector as a known nuclear localization marker. Images were taken 4 days after tobacco leaf infiltration. (A) pK7RWG2 empty vector carrying the 35S:RFP construct; (B) DcABRE1:RFP; (C) DcABRE3:RFP; (D) DcABRE4:RFP. Red channel: images taken under the Cy3 filter. Green channel: images taken under the fluorescein isothiocyanate filter. Scale bar: 25 μm.

Article Snippet: The subcellular localization of these three D. carota AREB/ABF transcription factors was determined by means of a double transient transformation of tobacco leaves with vectors carrying the sequences of DcAREB1 , DcAREB3 , and DcAREB4 fused to RFP and the pCAMBIA1302 vector, which possesses the GFP gene directed by the constitutive promoter 35SCaMV, with nuclear GFP localization (Marker Gene Technologies, Inc.). shows that the three D. carota AREB/ABF transcription factors are localized in the nucleus.

Techniques: Transformation Assay, Plasmid Preparation, Marker, Construct

Journal: bioRxiv

Article Title: Suppressor of Cytokine Signaling 3 Derived Peptide as a Therapeutic for Inflammatory, and Oxidative Stress Induced Damage to the Retina

doi: 10.1101/2023.09.04.556227

Figure Lengend Snippet: A . ARPE-19 cells in serum free medium were treated with R9-SOCS3-KIR or the control peptide (20 μM) for 1 hr followed by treatment with C5a (50 ng/ml) for 4 hr. They were then co-transfected with a mixture of plasmids with NF-kB promoter driven firefly luciferase and another plasmid with thymidine kinase driven Renilla luciferase as an internal control and grown for 24 hrs in 1% FBS containing medium. Cell lysates, were harvested and used to measure relative luciferase units using a Dual luciferase kit from Promega. **, p = 0.001. Error bars indicae the standard deviation. Abbrev: K, R9-SOCS3-KIR, Ctrl: R9-SOCS3-KIR-scrambled peptide. B. R9-SOCS3-KIR suppressed C5a-induced nuclear translocation of NF-κB subunit p65. ARPE-19 cells were grown overnight in 8 well plates, placed in serum-free medium and treated with R9-SOCS3-KIR or its control peptide at 20 μM for 1 hr, followed by addition of C5a (50 ng/ml) for 30 min. Cells were stained with an antibody to p65, followed by staining with Cy-3 conjugated secondary antibody and DAPI, and imaged in a fluorescence microscope using the same exposure time and light intensity. Scale bars equal 50 μm.

Article Snippet: The plasmid, pNF-kB-Luc that has the NF-kB promoter linked to firefly luciferase and a plasmid that constitutively expresses Renilla luciferase from the thymidine kinase promoter (pRL-TK-Luc) were purchased from Promega (Madison, WI). pRL-TK-Luc serves as an internal control to test the efficiency of transfection.

Techniques: Transfection, Luciferase, Plasmid Preparation, Standard Deviation, Translocation Assay, Staining, Fluorescence, Microscopy