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Fig. 5 | BMC Biology

Fig. 5

From: Intramembrane proteolysis of an extracellular serine protease, epithin/PRSS14, enables its intracellular nuclear function

Fig. 5

EICD-induced gene expression. a DEG analysis was performed between RNA samples from GFP- and GFP plus EICD-transfected cells. The results were visualized as MA plots in which log2 fold change (y-axis) and mean of normalized counts (x-axis) from FPKM (fragments per kilobase million) values are plotted. Genes with p values less than 0.05 are shown as red dots. Representative genes are indicated with arrows. b Specific antibodies against various cytokines (Additional file 1: Figure S3) were spotted onto the array in duplicate. Conditioned media from SP2bKD-10 cells (SP2bKD, upper) and EICD-expressing SP2bKD-10 cells (SP2bKD/EICD, lower) were incubated on the antibody array. Spots with more than 1.2-fold increases in media from SP2bKD/EICD cells are boxed in red. Positions of representative cytokines were indicated with arrows. c Cytokines showing more than a 1.2-fold increase in SP2bKD/EICD cells are shown. Cytokines increased in both the RNA-seq analysis and cytokine array are indicated with red font. d The 427 cells were transiently transfected with the GFP gene under the UAS promoter, GAL4 DNA binding domain (GAL4-DBD)-fused EICD, and tdTomato as a transfection marker. The degree of GFP expression in tdTomato-positive cells was analyzed using flow cytometry. The degree of GFP expression in each condition is shown as a bar graph (n = 3). Error bars indicate SEM. e 427 cells were transfected with GFP gene under CCL20 promoter, EICD, and FLAG-tagged EICD (or FLAG-tagged GAL4-DBD as a control), and tdTomato. The degree of GFP expression of tdTomato positive cells in each condition was subtracted by that of empty vector-transfected cells followed by normalization against that of 0.5 μg FLAG-EICD-transfected cells (left). Expression level of FLAG constructs are shown (right)

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