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Fig. 2 | Cell Communication and Signaling

Fig. 2

From: Non-POU Domain-Containing Octomer-Binding (NONO) protein expression and stability promotes the tumorigenicity and activation of Akt/MAPK/β-catenin pathways in human breast cancer cells

Fig. 2

C-terminal thr-pro motifs in NONO specifically interact with the WW domain of PIN1. a Yeast two-hybrid analysis shows the protein-protein interactions of NONO with the PIN1 and PIN1-WW domain. The yeast strains containing bait and prey plasmids were mated. The diploids were selected on double-dropout medium –LT (SD-Leu/-Trp) and quadra-dropout medium –LTHA (SD/-Leu/-Trp/-Ade/-His) for yeast two-hybrid screening. P53 BD/T antigen AD and Lam BD/T antigen AD was used as positive and negative controls. b Detection of FRET by acceptor photobleaching. HEK293T cells were co-transfected with PIN1-CFP/NONO-YFP, PIN-WW-CFP/NONO-YFP, PIN1-PPIase-CFP/NONO-YFP. After 48 h of transfection, the cells were fixed, the images of CFP and YFP were acquired using CFP and YFP channels, and the intensities were measured before and after acceptor photobleaching. Scale bar 25 µm. The bar graph depicting the FRET efficiency of various cyan/yellow pairs. Data are presented as mean ± standard deviation with n ≥ 3. p* < 0.05. c Presence of four the pro motifs in NONO ORF (CCDS14410.1). Source: National Canter for Biotechnology Information (https://www.ncbi.nlm.nih.gov/). d Site-directed mutagenesis identifies the thr-pro motifs essential for the interaction of NONO with the PIN1 WW domain. The effect of substituting threonine with glycine at thr-pro motifs within the NONO ORF was analyzed by yeast two-hybrid screening. The two thr-pro motifs at the C-terminus of NONO are crucial for the interaction of NONO with the PIN1 WW domain

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Cell Communication and Signaling

ISSN: 1478-811X