Fig. 4

The mutation in HaRxL103^Hind2 affects in planta subcellular localization. a Subcellular localization of GFP-HaRxL103^Emoy2 (GFP-103^Emoy2), GFP-HaRxL103^Hind2, and NLS- or nls-fused GFP-HaRxL103^Hind2. GFP-tagged HaRxL103^Emoy2 and HaRxL103^Hind2 variants were transiently co-expressed with RFP-HaRxL44^Emoy2 via agroinfiltration in N. benthamiana. The upper image is from the GFP channel, the middle image is from the RFP channel, and the lower image is the overlay of the GFP and RFP channels. Fluorescence intensity profile (GFP, green; RFP, red) across the white arrow was performed using the analyzing software (Leica, bottom). Scale bars, 10 μm. b HR cell death phenotypes when co-expressed of HaRxL103^Emoy2 and HaRxL103^Hind2 variants with RPP4 in N. benthamiana. The leaves inoculated with Agrobacterium containing the indicated gene constructs were photographed under UV at 3 dpi. c A predicted NLS sequence from HaRxL103^Emoy2 and HaRxL103^Hind2. Prediction of NLS in HaRxL103^Emoy2 or HaRxL103^Hind2 was done in cNLS Mapper. Position of predicted NLS and score for the prediction are indicated. Identical sequences are indicated in white on black. d Subcellular localization of GFP-fused predicted NLS from HaRxL103^Emoy2 (NLS^Emoy2) and HaRxL103^Hind2 (NLS^Hind2). GFP, GFP-NLS^Emoy2, and GFP-NLS^Hind2 were transiently co-expressed with RFP-HaRxL44^Emoy2 via agroinfiltration in N. benthamiana. Imaging was performed as described in (a). Scale bars, 10 μm