Journal: The Febs Journal

Article Title: The mitochondrial protein Sideroflexin 3 (SFXN3) influences neurodegeneration pathways in vivo

doi: 10.1111/febs.16377

Figure Lengend Snippet: Proteomics analysis and immunoblot validation of Sfxn3 ‐KO synaptosomes. (A) Scatterplot of the fold change ( Sfxn3 ‐KO/WT) distribution of 6,186 proteins identified by ≥ 2 unique peptides in proteomics screen of 1‐year‐old WT and Sfxn3 ‐KO mice brain synaptosomes. Dotted blue lines show the 20% up‐ and downregulated threshold and the specific data points beyond the threshold are indicated in red. (B) Top 10 canonical pathways identified from proteins altered by more than 20% in Sfxn3 ‐KO mice, as identified by Ingenuity Pathway Analysis. For complete proteomics dataset, see DataShare file: https://doi.org/10.7488/ds/3068 . (C) Immunoblot protein bands for CSPα and Uncleaved Caspase‐3 in synaptosome isolates from 1‐year‐old WT and Sfxn3 ‐KO mice brains. (D) Quantification of CSPα ( P = 0.026) and Uncleaved Caspase‐3 ( P = 0.017) protein levels in synaptosomes of 1‐year‐old WT and Sfxn3 ‐KO mice brains. * P < 0.05 in Mann‐Whitney U (CSPα) and in two‐tailed unpaired t ‐test (Uncleaved Caspase‐3). Quantitative data are shown as mean ± SEM ( N = 6 mice per genotype). Data values from two technical replicates were combined to produce the resulting mean. See also Fig. for proteomics experimental design scheme.

Article Snippet: After rinsing off the REVERT Total Protein Stain, the membrane was placed in Odyssey blocking buffer (Licor) for 30 min and then incubated with the following primary antibodies overnight at 4 °C: CSPα (Enzo, Farmingdale, NY, USA, Cat #: ADI‐VAP‐SV003‐E, 1 : 2000), Uncleaved Caspase‐3 (Cell Signaling Technologies, Danvers, MA, USA, Cat #: 9662, 1 : 1000), Cytochrome C (Abcam, Cambridge, UK, Cat #: ab110325, 1 : 1000), OXPHOS cocktail (Abcam Cat #: ab110411, 1 : 1000), Rieske Fe‐S (A‐5) (Santa Cruz Biotechnology, Dallas, TX, USA, Cat #: sc‐271609, 1 : 1000), SHMT2 (Atlas Antibodies Cat #: HPA020549, 1 : 750), CHCHD3 (Proteintech Cat #: 25625‐1‐AP, 1 : 2000), Bcl‐2 (Proteintech Cat #: 12789‐1‐AP, 1 : 1000), Mitofusin 1 (Proteintech Cat #: 13798‐1‐AP, 1 : 2000) and NDUFS1 (Abcam Cat #: ab169540, 1 : 2000).

Techniques: Western Blot, MANN-WHITNEY, Two Tailed Test

Journal: Scientific Reports

Article Title: Peripheral serotonin regulates postoperative intra-abdominal adhesion formation in mice

doi: 10.1038/s41598-017-10582-w

Figure Lengend Snippet: Serotonin accelerated angiogenesis in postoperative intra-abdominal adhesions by up-regulating VEGF expression. Postoperative intra-abdominal adhesion formation was induced by caecum rubbing, and the mice were sacrificed on the 7 th day after the operation. The adhesive tissues were harvested to detect the angiogenesis. ( A,B ) Immunohistochemistry for evaluating the levels of CD34 and VEGF in the adhesive tissues (magnification ×200); n = 6, mean ± SEM, * P < 0.05 vs. the WT group.

Article Snippet: The primary antibodies anti- CD34 antibody (bs-9752R, BIOSS, China, 1:300 dilution), anti- VEGF antibody (bs-0279R, BIOSS, China, 1:300 dilution), anti-TGF-β1 antibody (sc-146, Santa Cruz Biotechnology, USA, 1:400 dilution), anti-COX-2 antibody (bs-10411R, BIOSS, China, 1:300 dilution), anti-TIMP-1 antibody (bs-4600R, BIOSS, China, 1:300 dilution) and anti-MMP-2 antibody (bs-4599R, BIOSS, China, 1:300 dilution) were incubated overnight at 4 °C, and a secondary antibody was incubated for 1 h at room temperature.

Techniques: Expressing, Immunohistochemistry

Journal: British journal of cancer

Article Title: Expression of COX-2 and PGE synthase and synthesis of PGE(2)in endometrial adenocarcinoma: a possible autocrine/paracrine regulation of neoplastic cell function via EP2/EP4 receptors.

doi: 10.1054/bjoc.2001.2033

Figure Lengend Snippet: Figure 4 COX-2 (A), mPGES (B), PGE2 (C) and EP4 (D) are detected in endothelial cells of all carcinoma tissues. Vascular endothelial cells in endometrial adenocarcinoma were localised using antibodies raised against the human CD34 endothelial cell marker (E). The inset in (E) is a section that was stained with non-immune goal serum (CD34 negative control). Negative controls for the other antibodies are presented in Figures 2, 3 and 5B. Scale bar is 50 µm

Article Snippet: Subsequently the tissue sections were incubated with polyclonal goat anti-COX-2 antibody (sc-1745; Autogenbioclear, Wilts, UK) at a dilution of 1:400, polyclonal rabbit anti-mPGES antibody (catalogue number 160140; Cayman Chemical, Alexis Corporation-Europe, Nottingham, UK) at a dilution of 1:50, polyclonal rabbit anti-EP4 receptor (catalogue number 101770; Cayman Chemical) at a dilution of 1:500, polyclonal rabbit antiPGE2 antibody (kindly supplied by Professor RW Kelly, MRC Human Reproductive Sciences Unit, Edinburgh, UK) at a dilution of 1:100 or monoclonal mouse anti-human CD34 primary antibody (mca-547; Serotec, Oxford, UK) at a dilution of 1:25 at 4 ̊C for 18 h. Control tissue was incubated with either 5% non-immune antisera (CD34), goat anti-COX-2 antibody pre-adsorbed to blocking peptide (sc-1745p; Autogenbioclear), rabbit anti-mPGES antibody pre-adsorbed to blocking peptide (catalogue number 360140: Cayman Chemical), rabbit anti-EP4 pre-adsorbed to blocking peptide (catalogue number 101780; Caman Chemical) or © 2001 Cancer Research Campaign COX-2 and PGE2 in endometrial adenocarcinoma 1025 rabbit anti-PGE2 antibody pre-adsorbed to 10-fold excess PGE2 (Sigma).

Techniques: Marker, Staining, Negative Control