Fig. 5: BcSSP2 suppresses NUP62 phase separation-mediated selective nucleocytoplasmic transport. | Nature Communications

Fig. 5: BcSSP2 suppresses NUP62 phase separation-mediated selective nucleocytoplasmic transport.

From: The pathogen effector BcSSP2 suppresses the NPC phase separation to facilitate Botrytis cinerea infection

Fig. 5

a, b BcSSP2 inhibits MBP-GFP8W translocation into NUP62 hydrogel. Time-lapse photos were taken since MBP-GFP8W was supplied. The representative photos (a) and the translocation speeds (b) of MBP-GFP8W in NUP62, NUP62+control and NUP62 + BcSSP2 hydrogels are shown. Data are mean ± SD (n = 23 for NUP62, n = 24 for NUP62+control, n = 25 for NUP62 + BcSSP2,). Scale bar: 100 μm. c, d BcSSP2 inhibits MPK3 translocation. Time-lapse photos were taken since GFP-MPK3 was supplied. Representative photos (c) and relative fluorescent intensity (d) of GFP-MPK3 in NUP62+control or NUP62 + BcSSP2 hydrogels with WT Arabidopsis (Col-3) crude extracts are shown (n = 3). Dotted circles mark the hydrogel edge. Scale bars: 500 μm. e–j BcSSP2 suppresses nuclear accumulation of MPK3. For (e–g) WT Arabidopsis plants infected by B05.10 or bcssp2-12 at 36 hpi were subjected to analysis of total MPK3 proteins (e), nuclear MPK3 proteins (f) and PDF1.2 transcripts (g). For (h–j), the iBcSSP2 plants were treated with estradiol (ES) or DMSO (Mock), and then inoculated with bcssp2-12 knockout strain for 36 h and subjected to analysis of total MPK3 proteins (h), nuclear MPK3 proteins (i) and PDF1.2 transcripts (j). The protein levels of MPK3 were quantified based on the relative intensity of protein bands while the max intensity was normalized to 1 (e, f, h and right panel of i). The bands of MPK3 in (f) and (i) are presented with a short or long exposure time. Data are mean ± SE (n = 4 in (e, f, h) and n = 5 in (i), or mean ± SD (n = 3 in (g, j)). PEPC (phosphoenolpyruvate carboxylase), LSU (large subunit of rubisco protein) and H3 (Histone H3) serve as loading controls. MPK3 and PEPC were run on the same gel with the membrane cut for different antibodies; LSU was stained after exposure in (e and h). MPK3 and H3 were run on the same gel with the membrane cut for different antibodies in (f and i). Statistical significance was determined by two-sided Student’s t test. Source data are provided as a Source Data file.

Back to article page