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. Author manuscript; available in PMC: 2022 Mar 22.
Published in final edited form as: Science. 2021 Jun 17;373(6553):420–425. doi: 10.1126/science.abg7917

Fig. 4. Negatively charged residues in the conserved RNL motif are required for Ca2+ influx and cell death.

Fig. 4.

(A) In planta phenotypes (2 dpi) of plant RNLs in the presence of the Ca2+ channel blockers LaCl3 (La) and GdCl3 (Gd). Accumulation of YFP-tagged RNL variants was verified by SDS-PAGE and-immunoblotting, in Nb adr1 nrg1. (B, C, and E) Time course RNL-induced Ca2+ influx measurements with GCaMP3-transgenic Nb. (D and F) Time course conductivity measurement depicting RNL-triggered cell death in Nb. (G to J) [Ca2+]i in HeLa cells expressing NRG1.1 DV and variants (G and H) or ADR1 and variants (I and J). Black arrows indicate the addition of CaCl2. Letters indicate statistical significance (ANOVA with post hoc Tukey, P < 0.05). YFP-tagged RNL protein expression in Nb and HeLa cells was verified (fig. S16).