Extended Data Fig. 2: Generation and characterization of Arabidopsis gbpl mutants.
From: A phase-separated nuclear GBPL circuit controls immunity in plants
a, Immunoblot of GBPL3 protein level using α-GBPL3 rabbit polyclonal antibody in wild type (WT) (Col-0) and four independent T1 transformants generated via CRISPR/Cas9 targeting GBPL3 (predicted molecular weight 122 kDa). Ponceau S staining of a non-specific band (nsb) serves as a loading control. Proteins extracted from five-week-old plants. Experiment performed once only due to the limit of tissue materials from sterile plants. b, PAM sequences of gRNAs and the consequences of CRISPR-Cas cleavages in T1 generation. Line 15 is biallelic and line 16 is heterozygous. c, Chromatograms of sanger sequencing of the mutation sites in CRISPR lines in b. d, Fresh weight of plants (mean ± SD) shown above (top-right graph the same as Fig. 1b for comparison with left). e, Gene expression of GBPL1 and GBPL2 in T-DNA mutants. f, Immunoblot (α-GBPL3) of GBPL3 and EGFP–GBPL3 levels in complemented plants. Ponceau S staining of RbcL serves as a loading control. g, Growth of Psm ES4326 (Psm) at day 3 of infection. Inoculum, bottom left. h, Hpa Noco2 sporulation in 2~3-week-old plants at day 7. Inoculum, bottom left. i, GBPL proteins are not required for Pst DC3000 Avr challenge using gbpl mutant lines. Growth of Pst-Avr at day 3 of infection. Inoculum, bottom left. g–i, Box = 25th and 75th percentiles; bars = min and max values. Statistical analysis, comparison of mean via one-way ANOVA test (Bonferroni post hoc correction). ns, not significant. Individual data points represent biologically independent samples (d, g–i).
