Journal: Cancer Science

Article Title: Angiotensin II subtype 1a receptor signaling in resident hepatic macrophages induces liver metastasis formation

doi: 10.1111/cas.13306

Figure Lengend Snippet: Angiotensin II (Ang II )/ AT 1a axis enhances transforming growth factor‐β1 ( TGF ‐β1) expression in Kupffer cells. (a) Accumulation of F4/80 + cells, TGF ‐β1 + cells, and F4/80 + / TGF ‐β1 + cells in metastatic areas from WT and AT 1a KO mice on day 14. Double‐staining of liver sections with antibodies against F4/80 (red) and TGF ‐β1 (green) in WT and AT 1a KO mice. Expression of TGF ‐β1 was colocalized with F4/80 + cells. Metastatic area is delineated with the white dashed line. T, tumor. Scale bar = 100 μm. (b–d) Numbers of F4/80 + cells (b), TGF ‐β1 + cells (c), and F4/80 + TGF ‐β1 + cells (d) in metastatic areas from WT and AT 1a KO mice. Data are expressed as the means ± SD of six mice per group. * P < 0.05. (e) Expression of AT 1a and AT 1b in KUP 5 Kupffer cells. Data are expressed as the means ± SD of six mice per group. (f) Expression of TGF ‐β1 on KUP 5 cells under stimulation with Ang II . Expression of TGF ‐β1 was enhanced 6 h after stimulation with Ang II compared with control. There was no significant difference at 12 h. Data are expressed as the means ± SD of six mice per group. * P < 0.05 versus control.

Article Snippet: For the latter, sections were activated using Histo VT One (Nacalai Tesque, Yokohama, Japan) and then incubated overnight at 4°C with one of the following primary antibodies: (a) anti‐mouse F4/80 antibody (1:200, rat monoclonal, sc52664; Santa Cruz Biotechnology, Dallas, TX, USA); (b) anti‐mouse TGF‐β1 antibody (1:200, rabbit polyclonal, ab92486; Abcam, Cambridge, UK); (c) anti‐mouse GFP antibody (1:200, rabbit polyclonal, ab290; Abcam); (d) anti‐mouse Ang II type 1A receptor antibody (1:100, rabbit polyclonal, bs‐2132R; Bioss, Boston, MA, USA); (e) anti‐mouse desmin antibody (1:100, goat polyclonal, ab80503; Abcam); (f) anti‐mouse type I collagen antibody (1:100, rabbit polyclonal, ab21286; Abcam); or (g) anti‐mouse CD31 antibody (1:200, rabbit polyclonal, ab28364; Abcam).

Techniques: Expressing, Double Staining

Journal: Hypertension

Article Title: Identification of Bona Fide Alternative Renin Transcripts Expressed Along Cortical Tubules and Potential Roles in Promoting Insulin Resistance In Vivo Without Significant Plasma Renin Activity Elevation

doi: 10.1161/hypertensionaha.114.03394

Figure Lengend Snippet: Figure 7. A, The results of quantitative reverse transcription polymerase chain reaction analyses of inflammatory molecules in heart, kidney, and liver of both ARen2TG and wild-type littermates are shown as relative abundance, using GAPDH expression as an internal control. *, **, #, ##Significantly elevated compared with the expressions of wild-type littermates. Repeated measure ANOVA and subsequent post hoc analyses were performed. B, Representative results of in situ histological examinations for livers of both transgenic mice and wild-type littermate using rabbit polyclonal anti-angiotensin II antibodies (NBP1-30027; Novus Biologicals, LLC, Littleton, CO) were shown. a–c, Enhanced immunohistochemical staining in livers of transgenic mice compared with those of wild-type littermates (Bd), suggesting enhanced angiotensin II production in the organs.

Article Snippet: We performed experiments to compare ARen2TG mice with wild-type littermates in terms of following phenotypes: measurement of blood pressure,16,17 measurement of plasma renin activity,16,18,19 the results of in vivo near-infrared imaging using the renin substrate analog ReninSense 680 FAST (Perkin Elmer, Waltham, MA),20 the results of insulin tolerance tests,21 fasting plasma glucose levels, the expressions of inflammatory molecules in various tissues on quantitative RT-PCR analyses,16,18,19 and in situ histological examinations for livers of both transgenic mice and wild-type littermate using rabbit polyclonal anti-angiotensin II antibodies (NBP1-30027; Novus Biologicals, LLC, Littleton, CO).22,23 All experiments were performed according to the protocols of manufacturer, the references, and our reports as specified above.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Expressing, Control, In Situ, Transgenic Assay, Immunohistochemical staining, Staining

Journal: Molecular Cell

Article Title: Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

doi: 10.1016/j.molcel.2021.07.013

Figure Lengend Snippet:

Article Snippet: Primary antibodies used were anti-β-catenin (# 610153, BD Transduction Laboratories), anti-phospho-β-catenin pS33, pS37, pT41 (# 9561, Cell Signaling Technology), anti-non-phospho-β-catenin S33, S37, T41 (#8814, Cell Signaling Technology), anti-phospho-β-catenin pS45 (# 9564, Cell Signaling Technology), anti-StrepMAB-Classic (# 2-1507-001, IBA Life Sciences) to detect dStrepII-AXIN1, anti-AXIN1 (# 2087, Cell Signaling Technology), anti-APC (# NB100-667, Novus Biologicals), anti-GSK3β (# 12456, Cell Signaling Technology), anti-phospho-GSK3β pS9 (# 5558, Cell Signaling Technology), anti-ubiquitin K48 linkage-specific (# ab140601, Abcam), anti-ubiquitin K63 linkage-specific (# ab179434, Abcam), anti-ubiquitin P4D1 clone (# BML-PW0930, Enzo Lifesciences).

Techniques: Virus, Recombinant, Protease Inhibitor, Ubiquitin Proteomics, Silver Staining, Staining, Plasmid Preparation, Mutagenesis, Software, Multi-Angle Light Scattering, Fluorescence