Journal: Scientific Reports

Article Title: Phytocyanin-encoding genes confer enhanced ozone tolerance in Arabidopsis thaliana

doi: 10.1038/s41598-022-25706-0

Figure Lengend Snippet: Cellular localisation and the ability of AtUC5-GFP fusion proteins to confer O 3 tolerance. ( a ) Diagrams depicting the chimeric genes to produce fusion proteins in transgenic plants. ( b ) Results of western blotting to detect fusion protein expression in extracts of transgenic plants. Equal amounts (3 µg) of protein extracted from plants of various lines were subjected to electrophoresis and probed with anti-GFP antibody using Simple Western™ (Bio-Techne, Minneapolis, USA). GFP was added to the protein extract from the wild-type plant just before the electrophoresis for the lane “wt + GFP.” Results were obtained as digital data for each sample and exhibited as band images as their full-length blots. ( c ) Microscopic observation of GFP fluorescence in epidermal tissues of transgenic plants. α fluorescence images of epidermal tissue of a leaf transgenic for 35S:GFP-AtUC5 . β fluorescence image of epidermal tissue of a leaf transgenic for 35S:AtUC5-GFP . γ fluorescence image of epidermal tissue of a leaf transgenic for 35S:GFP-AtUC5 after treatment with hyperosmotic solution. δ fluorescence image of the epidermal tissue of a leaf transgenic for 35S:AtUC5-GFP after treatment with hyperosmotic solution. Arrowheads and arrows denote stomata and apoplastic matrices, respectively. ( d ) Diagrams depicting the presumed structures and localizations of fusion proteins in transgenic plants. In 35S:GFP-AtUC5 plants, the fusion protein is released from the plasma membrane after digestion at the GPI moiety by phospholipases. ( e ) Images of various lines 1 day after 2 h exposure to fresh air or 0.3 µL L −1 O 3 under 420 µmol photons m −2 s −1 illumination. Wt wild-type plants, GFP green fluorescent protein, 35S:GFP-AtUC5 35S:AtUC5-GFP , transgenic plants producing fusion proteins, 35S cauliflower mosaic virus 35S promoter, N a fragment encoding the N-terminal signal peptide of AtUC5, PLD plastocyanin-like domain, ALR arabinogalactan protein-like region, GPI glycosylphosphatidylinositol moiety, CGAS C-terminal glycosylphosphatidylinositol-anchored signal, FA fresh air .

Article Snippet: Equal amounts (3 µg) of protein extracted from plants of various lines were subjected to electrophoresis and probed with anti-GFP antibody using Simple Western™ (Bio-Techne, Minneapolis, USA).

Techniques: Transgenic Assay, Western Blot, Expressing, Electrophoresis, Fluorescence, Membrane, Virus

Journal: eLife

Article Title: A vibrissa pathway that activates the limbic system

doi: 10.7554/eLife.72096

Figure Lengend Snippet: ( A ) Viral method used for labeling paralemniscal projections. ( B ) Labeling of interpolaris cells after injection of G-pseudotyped Lenti-Cre virus in Po thalamus, and a Cre-dependent AAV that expresses GFP in the vibrissa-responsive sector of SpVIr. Horizontal section. ( C ) Anterograde labeling of terminal fields in Po thalamus and zona incerta. Sagittal section. ( D ) Anterograde labeling in the KF/PBc. Horizontal section. ( E ) Anterograde labeling in the ITr. Horizontal section. ( F ) Anterograde labeling in the dorsal sector of the facial nucleus. Horizontal section. ( G ) Anterograde labeling in the MdD and cervical cord. Horizontal section. ( H ) Population peristimulus time histogram of spike discharges evoked in KF (22 cells) by air puff deflection of the vibrissae in the anesthetized rat. A representative response is shown in the insert. ( I ) Example of a vibrissa responsive KF cell labeled by juxtacellular delivery of Neurobiotin. Horizontal section. ( J ) Location of eight juxtacellularly labeled KF cells. Horizontal brainstem sections in ( I ) and ( J ) were counterstained for cytochrome oxidase. Horizontal section. ( K ) Spectral coherence of spontaneous discharges of KF cells with the respiratory cycle at the respiratory frequency; 1–3 Hz. Note that, in contrast with the respiratory units (blue dots), spontaneous discharges of vibrissa-responsive cells (red triangles) display low coherence with respiration. ( L ) Population peristimulus time histogram of spike discharges evoked in MdD (33 cells) by air puff deflection of the vibrissae. ( M ) Recording site in the MdD labeled by an iontophoretic injection of Chicago Sky Blue. This coronal section was counterstained for cytochrome oxidase and a negative image was generated. Coronal section. See for additional anatomical data. Abbreviations for all anatomy: 5n, root of the trigeminal motor nucleus; 5N, trigeminal motor nucleus; 5t, trigeminal tract; 7n, facial nerve tract; 7N, facial nucleus; Amb, ambiguus nucleus; APT, anterior pretectal nucleus; BSTL, bed nucleus of the stria terminalis; CeA, central amygdala; Cerv Cord, cervical cord; CM/PC, central medial/paracentral thalamic nuclei; CPu, caudate putamen; DR, dorsal raphe; EW, Edinger-Westphal; Hab, habenula; IML, intermedio-lateral column of the spinal cord; ITr, intertrigeminal region; KF, Kölliker-Fuse nucleus; KF/PBc, Kölliker-Fuse/parabrachial complex; mcp, middle cerebellar peduncle; MdD, dorsal part of the medullary reticular formation; MdV, ventral part of the medullary reticular formation; mt, mammillothalamic tract; NA, nucleus ammbiguus; NTS, nucleus of the solitary tract; opt, optic tract; PAG, periaqueductal gray; PB, parabrachial nuclei; PC, paracentral thalamic nucleus; PCRt, parvicellular reticular formation; PLH, posterior lateral hypothalamus; Po, posterior nuclear group of the thalamus; PrV, principal trigeminal nucleus; RN, red nucleus; Rt, reticular thalamic nucleus; s5, sensory root of the trigeminal nerve; SC, superior colliculus; scp, superior cerebellar peduncle; SpVC, caudalis division of the spinal trigeminal complex; Sol, nucleus of the solitary tract; SpVIc, caudal sector of the interpolaris trigeminal nucleus; SpVIr, rostral division of the interpolaris nucleus; TG, trigeminal ganglion; VLL, ventral nucleus of the lateral lemniscus; VPL, ventral posterolateral thalamic nucleus; VPM, ventral posterior medial nucleus; VPCc, parvicellular sector of the ventral posteromedial thalamic nucleus; VPPc, parvocellular part of the ventral posterior thalamic nucleus; VRG, ventral respiratory group; vsc, ventral spinocerebellar tract; ZIv, ventral division of zona incerta.

Article Snippet: For brightfield microscopy, sections were first counterstained for cytochrome oxidase (MilliporeSigma), and then immunoreacted with a rabbit anti-GFP antibody (1:1000; Novus Biological), a biotinylated horse anti-rabbit IgG (1:200; Vector Labs), the avidin/biotin complex (Vectastain ABC Kit; Vector Labs), and the SG peroxidase substrate (ImmPACT SG Substrate; Vector Labs).

Techniques: Labeling, Injection, Virus, Generated

Journal: eLife

Article Title: A vibrissa pathway that activates the limbic system

doi: 10.7554/eLife.72096

Figure Lengend Snippet:

Article Snippet: For brightfield microscopy, sections were first counterstained for cytochrome oxidase (MilliporeSigma), and then immunoreacted with a rabbit anti-GFP antibody (1:1000; Novus Biological), a biotinylated horse anti-rabbit IgG (1:200; Vector Labs), the avidin/biotin complex (Vectastain ABC Kit; Vector Labs), and the SG peroxidase substrate (ImmPACT SG Substrate; Vector Labs).

Techniques: Plasmid Preparation, Recombinant, Virus, Software