Fig. 3
From: Cell entry of a host-targeting protein of oomycetes requires gp96
SpHtp3 self-translocates into host cells via its C-terminus. a Self-translocation of SpHtp3-mRFP wt, SpHtp3-mRFP21–55 (containing the RTLR sequence), a mutant of SpHtp3-mRFPRTLR/GTLG and mRFP only into living RTG-2 cells at pH 5.5. Scale bar: 20 µm (n = 3). b Quantitative FACS analysis of RTG-2 cells from Figs. 2d and 3a. Error bars denote s.e.m. (n = 3). *** p < 0.001 (one way ANOVA). c Homology model of SpHtp3 calculated by YASARA containing the natural ligands (nucleotide analogue and Ca2+ ion (yellow)). The inset shows the structural details of the very C-terminus of SpHtp3 containing a short charged helix built up by an YKARK sequence. d Self-translocation of a FITC-labelled C-terminal peptide of SpHtp3 (201–211 aa, P) into RTG-2 cells at pH 5.5. Nuclei are indicated by dashed lines. Error bars denote s.e.m. (cells: 50). ***p < 0.001 (t-test). Scale bar: 20 µm (n = 3). e Self-translocation of a mutant of SpHtp3-mRFP (K208A/R210A, KR) in comparison to SpHtp3-mRFP wt into RTG-2 cells at pH 5.5. Nuclei are indicated by dashed lines. Error bars denote s.e.m. (cells: 50). ***p < 0.001 (t-test). Scale bar: 20 µm (n = 3)
