Journal: PLoS Pathogens

Article Title: Functionally Redundant RXLR Effectors from Phytophthora infestans Act at Different Steps to Suppress Early flg22-Triggered Immunity

doi: 10.1371/journal.ppat.1004057

Figure Lengend Snippet: ( A ) Confocal microscopy of A. thaliana protoplasts expressing GFP-SFI1 or myr-GFP-SFI1 12 h after transfection. Representative optical sections of bright field and merged GFP (in green) and chloroplast (in blue) fluorescence images are shown. ( B ) Representative confocal microscope images of N. benthamiana cells expressing GFP-SFI1 and myr-GFP-SFI1 (left panels, in green) with the nucleolar marker RFP-fibrillarin (right panels, in red); the merged images are shown in the central panels. ( C ) Measurement of pFRK1-Luc reporter activity in A. thaliana protoplasts 6 h after flg22 treatment in the presence of GFP (control), AvrPto, GFP-SFI1 or myr-GFP-SFI1. Pooled data from four experiments are presented as mean ± SEM. Significant differences (p<0.05) in luciferase activity (denoted *) were determined using one-way ANOVA followed by Dunnett's multiple comparison test. ( D, E ) Effect of transient expression of GFP-SFI1 and myr-GFP-SFI1 on P. infestans colonization of N. benthamiana . Mean lesion diameter ( D ) and typical disease symptoms ( E ) are shown for P. infestans 7 days post-inoculation over sites on leaves where an effector construct was agro-infiltrated 1 day earlier. * represents statistical significance (p<0.001) using one-way ANOVA. ( F ) Immunoblot using anti-GFP antibody showing that both GFP-SFI1 and myr-GFP-SFI1 are stable and intact fusion proteins (arrowed) in planta .

Article Snippet: The expression of GFP-tagged PiRXLR effectors was assessed by immunoblotting using polyclonal anti-GFP antibody produced in rabbit or in goat (Acris Antibodies) at a 1/3000 dilution in 5% BSA in TBS-T. For this, protoplast samples were collected 12 (for S. lycopersicum ) or 24 h (for A. thaliana ) after transfection and SDS-PAGE and immunoblotting were carried out as described above.

Techniques: Confocal Microscopy, Expressing, Transfection, Fluorescence, Microscopy, Marker, Activity Assay, Luciferase, Construct, Western Blot

Journal: Cell Reports Methods

Article Title: NeuroGT: A brain atlas of neurogenic tagging CreER drivers for birthdate-based classification and manipulation of mouse neurons

doi: 10.1016/j.crmeth.2021.100012

Figure Lengend Snippet:

Article Snippet: The sections were washed with 10 mM Tris-HCl (pH 7.4), 130 mM NaCl and 0.1% Tween 20 (TBST) and incubated with rabbit anti-GFP antibody (1:1000, #598, MBL International Cat# 598, RRID: AB_591816 ) diluted in PBS containing 0.1% Tween 20 and 0.5% blocking reagent (FP1020, PerkinElmer) overnight at 4°C.

Techniques: Recombinant, Software