Extended Data Fig. 4 | RHA3A/B interacts with BIK1 in vivo. a, BIK1 interacts
with RHA3A in a co-IP assay. RHA3A–HA was co-expressed with BIK1–FLAG or
control in protoplasts and then treated with 100 nM f lg22 for 15 min. Left, the
co-IP assay was carried out with anti-FLAG agarose and immunoprecipitated
proteins were immunoblotted with anti-HA or anti-FLAG antibody. Right,
BIK1–FLAG and RHA3A–HA proteins. b, RHA3A expression (mean ± s.e.m.) in
pRHA3A::RHA3A-FLAG/pBIK1::BIK1-HA transgenic plants. qRT–PCR was carried
out to detect RHA3A transcripts using ACTIN2 as a control. Relative gene
expression in wild-type (set as 1), pBIK1::BIK1-HA (Ctrl) and two independent
transgenic lines (lines 7 and 10) is shown. One-way ANOVA, n = 3. c, BIK1
associates with RHA3B independent of f lg22 treatment. RHA3B–HA was
co-expressed with BIK1–FLAG or control in protoplasts and then treated with
100 nM f lg22 for 15 min. Left, co-IP assay was carried out with anti-FLAG
agarose and immunoprecipitated proteins were immunoblotted with anti-HA
or anti-FLAG antibody. Right, BIK1–FLAG and RHA3B–HA proteins before
immunoprecipitation. d, FLS2 interacts with RHA3A and RHA3B in a co-IP
assay. Experiments were repeated three times with similar results.