Fig. 2: Combination of RNA-Seq and proteomics revealed transcriptional-translational inconsistency of TRI cluster genes under FgSfp1 deprivation.
a Wild-type PH-1 strain bearing FgSfp1-GFP constructs were incubated in yeast extract peptone dextrose medium (YEPD) at 28°C, 180 rpm for 24 h. The GFP fluorescent signal was detected by confocal laser scanning microscope (both). The nucleus of F. graminearum was co-stained with nuclear staining 4′-6-diamidino-2-phenylinodle (DAPI) and the images were captured at the same field with GFP signal channel (middle lanes). Scale bar = 40 μm. Fluorescence intensity and colocalization analysis by using imageJ software (line scan plots) represent fluorescent signals distributed in both nucleus and cytoplasm area. Horizontal axis indicates the distance. Results are from three independent experiments. b, c RNA-Seq data shows that the individual transcript levels of TRI genes are differentially expressed in wild-type PH-1 and ∆FgSfp1 strains. Depicted by bar plots (FPKM: Fragments Per Kilobase of exon model per Million mapped fragments). The numeric values are log2(FPKM + 1). d Relative mRNA expression levels of TRI5, TRI1, TRI101 in the ∆FgSfp1 mutant compared with wild-type PH-1, detected by qRT-PCR assay. Line bars in each column represent mean ± s.d. (n = 3). FgTUBULIN (locus tag FGSG_09530) was used as an internal loading control. e Relative Tri protein expression levels quantified by proteomics analysis. The results represented by bar plot. Tri protein relative abundance decreased in the ∆FgSfp1 mutant (red) compared with the wild-type PH-1 strain (blue). The data was analyzed with MaxQuant. Line bars in each column represent mean ± s.d. (n = 9). Relative Tri protein expression levels quantified by proteomics analysis. The results represented by bar plot. Tri protein relative abundance decreased in the ∆FgSfp1 mutant (red) compared with the wild-type PH-1 strain (blue). The data was analyzed with MaxQuant. Line bars in each column represent mean ± s.d. (n = 9).
