Journal: The Plant Pathology Journal

Article Title: Cucumber Mosaic Virus 1a Protein Interacts with the Tobacco SHE1 Transcription Factor and Partitions between the Nucleus and the Tonoplast Membrane

doi: 10.5423/PPJ.FT.03.2021.0045

Figure Lengend Snippet: Pull-down assay for detection of CMV 1a:SHE1 interaction. Extracted Escherichia coli -expressed fusion proteins (SHE1-MBP, 1a-GST, and Fb2-GST), AP, for SHE1-MBP and associated proteins, using amylose beads, were fractionated by SDS-PAGE and either stained with CBB (lower panel) or subjected to western blotting and probing with antisera to GST (upper panel) or CMV 1a protein (middle panel). The GST-fusion with nucleolar protein fibrillarin (Fb2) and free GST were used as negative controls. The CBB stained proteins served as loading controls. CMV, cucumber mosaic virus; AP, affinity-purified; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; CBB, Coomassie Brilliant Blue; GST, glutathione S-transferase; Fb2, fibrillarin 2. AP expressed protein samples loaded onto the gel were as follows: lane 1, SHE1-MBP/Empty-GST; lane 2, SHE1-MBP/CMV 1a-GST; lane 3, SHE1-MBP/Fb2-GST; and lane 4, SHE1-MBP only. The mol. wt. markers are shown to the left of the upper and middle panels. Protein sizes were as follows: MBP, 42 kDa; GST, 26 kDa; Fb2, 39 kDa; SHE1, 26.5 kDa; and CMV 1a, 111 kDa. The doublet band of ca. 80 kDa present in the anti-CMV 1a probed blot is a non-specific cross-reacting protein.

Article Snippet: To assess purification, the eluted proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and were either visualized by Coomassie Brilliant Blue staining or transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA, USA) for western blot analyses with polyclonal anti-GST (Sigma-Aldrich).

Techniques: Pull Down Assay, SDS Page, Staining, Western Blot, Virus, Affinity Purification, Polyacrylamide Gel Electrophoresis

Journal: Scientific Reports

Article Title: (Pro)renin receptor is crucial for Wnt/β-catenin-dependent genesis of pancreatic ductal adenocarcinoma

doi: 10.1038/srep08854

Figure Lengend Snippet: (a) Plasma s(P)RR levels in 20 patients with PDAC and 20 healthy matched controls. Data are mean ± SD values. The boxes encompass the twenty-fifth through seventy-fifth percentiles of results. The lines through the middle of each box represent the medians. The maximum and minimum values within 1.5 × interquartile range (IQR) are shown as whisker caps. Average values are indicated by dots in the boxes. (b) s(P)RR in conditioned medium was assayed for HPDE cells and six different PDAC cell lines. Consistent results were observed when three experiments were repeated. Loading control was determined by CBB staining.

Article Snippet: To obtain a loading control of s(P)RR expression, we used the CBB (Coomassie Brilliant Blue) staining solution (Wako, Osaka, Japan, catalog #17400553) to visualize the protein bands.

Techniques: Whisker Assay, Staining