Journal: Neurobiology of aging

Article Title: Prostate stem cell antigen interacts with nicotinic acetylcholine receptors and is affected in Alzheimer's disease.

doi: 10.1016/j.neurobiolaging.2015.01.001

Figure Lengend Snippet: Fig. 1. Prostate stem cell antigen (PSCA) is soluble and co-purifies with the a4 nAChR subunit in human cortex. (A) The antiserum directed against PSCA recognizes the human recombinant GST-tagged PSCA protein (total molecular weight 34.2 kDa) from 1 to 16 ng/well at the expected band size. Endogenous PSCA from mouse and human cortical tissue (8 mg total protein/well) is detected at approximately 24 kDa, and the amount in the cortical samples was compared with the known concentrations of re- combinant PSCA protein. (B) Images of Western blot showing PSCA protein levels in cortical tissue from human (protein concentration 4, 8, and 12 mg/well) and mouse (8, 15, and 30 mg/well) in the absence (peptide) and the presence of PSCA peptide (þpeptide). (C) Representative images of Western blots showing PSCA and Lypd6 protein levels in soluble and membrane fractions of human temporal cortical tissue. The membrane receptor proteins b2 nAChR and GluR2 are used as control. (D) Mag- netic beads covalently coupled with PSCA recombinant protein were incubated with cortical homogenates from human temporal cortex followed by detection of nAChR subunits by Western blot. Homogenates before (input) and after affinity purification (output) as well as the negative control (CTRL) were loaded.

Article Snippet: Validation of PSCA antibody (1:1,000, Novus Biologicals) was done with co-incubation with the PSCA peptide, which had been used as immunogen (1:100, # NB100-91938PEP, Novus Biologicals).

Techniques: Recombinant, Molecular Weight, Western Blot, Protein Concentration, Membrane, Control, Incubation, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: Study of dietary‑induced progression of psoriasis‑like mice based on gut macrophage polarization.

doi: 10.3892/etm.2023.11976

Figure Lengend Snippet: Figure 5. Protein expressions of Jagged1/Notch1/HES5 and TLR‑2/IκB‑α/NF‑κB p65 signaling pathway on skin lesions of imiquimod‑induced psoriasis model. (A and a) Western blot analysis of Notch signaling and NRP1 in skin lesions. The mammalian Notch signaling pathway consists of four Notch receptors (Notch 1, 2, 3, and 4) and five Notch ligands (Jagged 1, and 2, and Delta‑like 1, 3 and 4). HES5 is an important downstream target gene of Notch signaling pathway. NRP1 is involved in keratinocyte hyperproliferation of psoriasis accompanied by inflammation activated angiogenesis. (B and b) Western blot analysis of TLR‑2/IκB‑α/NF‑κB p65 signaling in skin lesions. Values are expressed as mean ± SD (*P<0.05, **P<0.01 vs. the normal control; #P<0.05, ##P<0.01, ###P<0.001 vs. the psoriasis group) and were statistically analyzed with one‑way ANOVA. NC, normal control; P, psoriasis; P + SF, psoriasis + SF; SF, stimulating food; p‑, phosphorylated.

Article Snippet: Proteins were transferred onto a PVDF membrane (MilliporeSigma), and then incubated with primary antibodies specific for NF‐κB p65 (cat. no. 6956), phospho‐NF‐κB p65 (cat. no. 3033), Toll‐like receptor‐2 (cat. no. 2229S), IKBα (cat. no. 9242), Jagged‐1 (cat. no. 70109s), Notch‐1 (cat. no. 3608s; all CST; 1:1,000), Hes‐5 (cat. no. ab25374; Abcam; 1:1,000) and anti‐β‐actin (cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.; 1:2,000) overnight at 4 ̊C and then followed by incubation with horseradish peroxidase‐conjugated secondary anti‐mouse or anti‐rabbit IgG (cat. nos.

Techniques: Western Blot, Control