Journal: International journal of oncology

Article Title: USP22 promotes the G1/S phase transition by upregulating FoxM1 expression via β-catenin nuclear localization and is associated with poor prognosis in stage II pancreatic ductal adenocarcinoma.

doi: 10.3892/ijo.2014.2531

Figure Lengend Snippet: Figure 6. USP22 expression affects FoxM1 expression and Wnt/β-catenin pathway activation. (A) Triple IF staining for FoxM1 (red), β-catenin (green) and nuclei (DAPI, blue) was performed on PANC-1 cells that were transfected by control-siRNA, USP22-siRNA-1 (50 nM) or USP22-siRNA-2 (150 nM). (B) Triple IF staining for FoxM1 (red), β-catenin (green) and nuclei (DAPI, blue) was performed on CFPAC-1 cells that were transfected by vector USP22-1 (2 µg) or USP22-2 (10 µg). (C and D) RT-PCR and western blot analysis of FoxM1 and β-catenin in PANC-1 cells that were transfected by control- siRNA, USP22‑siRNA-1 (50 nM) or USP22-siRNA-2 (150 nM) (left panel) and CFPAC-1 cells that were transfected by vector, USP22-1 (2 µg) or USP22-2 (10 µg) (right panel). (E) Cellular levels of Axin2, c-Myc and LEF-1 in PANC-1 cells that were transfected by control-siRNA, USP22-siRNA-1 (50 nM) or USP22‑siRNA-2 (150 nM) (left panel) and CFPAC-1 cells that were transfected by vector, USP22-1 (2 µg) or USP22-2 (10 µg) (right panel). (F) Activities of TOP-Flash and FOP-Flash in PANC-1 cells that were transfected by control-siRNA, USP22-siRNA-1 (50 nM) or USP22-siRNA-2 (150 nM) (left panel) and CFPAC-1 cells that were transfected by vector, USP22-1 (2 µg) or USP22-2 (10 µg) (right panel). (G) Cytoplasmic and nuclear levels of FoxM1 and β-catenin in PANC-1 cells that were transfected by control-siRNA, USP22-siRNA-1 (50 nM) or USP22-siRNA-2 (150 nM). (H) Cytoplasmic and nuclear levels of FoxM1 and β-catenin in CFPAC-1 cells that were transfected by vector USP22-1 (2 µg) or USP22-2 (10 µg).

Article Snippet: Membranes were blocked in a buffer (TBS: 50 mM Tris-HCl, 150 mM NaCl, pH 7.4) containing 5% bovine serum albumin and 0.1% Tween-20, followed by incubation with the primary antibodys USP22 (ab4812, 1:2,000, Abcam), FoxM1 (SC-502, 1:200, Santa Cruz Biotechnology), cyclin D1 (60186-1-lg, 1:100), p21 (10355-1-AP, 1:100), p27 (10567-1-AP, 1:100), cdk4 (11026-1-AP, 1:500), cdk6 (14052-1-AP, 1:500, all from Proteintech, Chicago, IL, USA), β-catenin (SC-7963, 1:500, Santa Cruz Biotechnology), Axin2 (EPR2005-2, 1:2,000), LEF (EP2030Y, 1:2,000, both from Abcam), c-Myc (10057-1-AP, 1:200) and LMNB1 (12987-1-AP, 1:1,000) or β-actin (20536-1-AP, 1:2,000, all from Proteintech) diluted in the same buffer.

Techniques: Expressing, Activation Assay, Staining, Transfection, Control, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Materials Characterization

Article Title: Space rocks and optimising scanning electron channelling contrast

doi: 10.1016/j.matchar.2018.06.001

Figure Lengend Snippet: Fig. 3. Forescatter diode (FSD) imaging with varying detector tilt. (a) At low detector tilt, contrast is optimised to reveal surface topography, demonstrating black-white contrast (a-i and a-ii) either side of a preferentially polished region of the FCC phase, and objects are located on the top of the surface show the ‘RGB’ plume artefact due to preferential shadowing of electrons for each diode use to form the red, blue, and green diodes. As the detector is tilted towards higher screen positions as compared to the sample (b and c) the topography contrast is suppressed and the orientation contrast between grains and between sub-grains increases. Inserts are of higher magnification regions and examples of raw EBSD patterns are given to indicate the relative position of the detector and the incident electron signal. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Forescatter electron imaging was captured using the ARGUS imaging system, mounted on a Bruker e-Flash FS EBSD detector and EDS data was acquired using a Bruker XFlash 6|60 detector.

Techniques: Imaging

Journal: Materials Characterization

Article Title: Space rocks and optimising scanning electron channelling contrast

doi: 10.1016/j.matchar.2018.06.001

Figure Lengend Snippet: Fig. 4. Forescatter diode (FSD) imaging with varying detector distance. (a) The ‘near’ position has a detector approx. 15 mm from the sample, (b) ‘medium’ at 20 mm, and (c) ‘far’ at 25 mm. Contrast in the near condition is strongly dominated by topography, and the far condition (for this detector tilt) high- lights contrast for the crystal orientation. Inserts are of higher magnification regions and an example EBSD pattern to indicate the relative position of the detector and the incoming electron signal.

Article Snippet: Forescatter electron imaging was captured using the ARGUS imaging system, mounted on a Bruker e-Flash FS EBSD detector and EDS data was acquired using a Bruker XFlash 6|60 detector.

Techniques: Imaging

Journal: Scientific Reports

Article Title: Smurf1 promotes gastric cancer progression by regulating Axin2-dependent Wnt signaling pathway

doi: 10.1038/s41598-025-23707-3

Figure Lengend Snippet: Smurf1 promoted Axin2 ubiquitination and ubiquitin-dependent degradation. ( A ) qRT-PCR analysis of Axin1 and Axin2 mRNA levels in AGS cells after Smurf1 overexpression. ( B ) Western blot analysis of Axin1 and Axin2 protein levels in AGS cells after Smurf1 overexpression. ( C ) Co-IP assay of the direct interaction between Smurf1 and Axin2 in AGS cells using the Smurf1 antibody, followed by western blot analysis using the Axin2 antibody. ( D and E ) After Smurf1 overexpression, de novo protein synthesis was inhibited in AGS cells using 40 µM of Chx, and then Axin2 protein levels were measured using western blot assays at different time points. ( F ) After Smurf1 overexpression in AGS cells, a Co-IP assay was carried out using the Axin2 antibody, followed by western blot analysis using the ubiquitin antibody (Left: Smurf1 OE + Axin2 IP + Ub WB; Middle: no Smurf1 OE + Axin2 IP + Ub WB; Right: Smurf1 OE + IgG IP + Ub WB. OE: overexpression, Ub: ubiquitin, WB: western blot). ( G ) After Smurf1 overexpression and treatment with 20 µM of MG132 in AGS cells, Axin2 protein levels were measured using the western blot assay. The results were shown as the median (1st quartile and 3rd quartile). * p < 0.05.

Article Snippet: For the ubiquitination analysis, western blot was carried out to assess Axin2 protein in AGS cells following Smurf1 overexpression and MG132 (20 μM, MCE) treatment for 24 h.

Techniques: Ubiquitin Proteomics, Quantitative RT-PCR, Over Expression, Western Blot, Co-Immunoprecipitation Assay