Journal: Journal of Advanced Research

Article Title: Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53

doi: 10.1016/j.jare.2023.09.041

Figure Lengend Snippet: Antibody list.

Article Snippet: Membrane blocking was performed using 5 % skimmed milk in Tris-buffered saline for 1 h, followed by incubation with the appropriate primary antibodies against total ERK (Cat No. B7074, Assay Biotechnology, Fremont, CA, USA), phosphorylated ERK (Cat No. 9101 s, Cell Signaling Technology, Danvers, MA, USA), total AKT (Cat No. CSB-PA000855, CUSABIO, Houston, TX, USA), phosphorylated AKT (Cat No. CSB-PA008120, CUSABIO), IKKα (Cat No. 11930, Cell Signaling Technology), FGFR1 (Cat No. NBP2-33784, Novus Biologicals, Centennial, CO, USA), phosphorylated FGFR1 (Tyr653, Tyr654) (Cat No. 44-1140G, Invitrogen) polyclonal Antibody, FRS2 antibody (Cat No. 3836–100, BioVision, Waltham, MA, USA), phosphorylated FRS2-alpha (Cat No. 3864S, Cell Signaling) or ACTIN (Cat No. sc-8432, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4 °C.

Techniques: Staining, Plasmid Preparation

Journal: Frontiers in Molecular Neuroscience

Article Title: The Alpha Isoform of Heat Shock Protein 90 and the Co-chaperones p23 and Cdc37 Promote Opioid Anti-nociception in the Brain

doi: 10.3389/fnmol.2019.00294

Figure Lengend Snippet: Hsp90α regulates opioid signal transduction in the brain. Male CD-1 mice had vehicle or 0.1 nmol KUNA115 injected icv , 24 h, followed by vehicle or 0.1 nmol DAMGO icv , 10 min. Periaqueductal gray region analyzed by Western blot. All data reported as the mean ± SEM, with sample sizes of mice/group noted in each graph; all experiments performed in four technical replicates. For ERK, JNK and Akt, data analyzed by two-way ANOVA with Fisher’s Least Significant Difference post hoc test; **,***,**** p < 0.01, 0.001, 0.0001 vs. Vehicle:Vehicle group. For Hsp70 and STAT3 data analyzed by Unpaired 2-Tailed t -test; * p < 0.05 vs. same target Vehicle group. (A) Representative sample blots shown for each target, with MW indicated for each protein. Each pair of images for one target (e.g., p-Akt) were from the same blot, but discontinuous, so they are separated to denote this fact. (B) All Western data quantitated by target. ERK, JNK, and Akt are phosphorylated protein signal normalized to total protein. Hsp70 and STAT3 are normalized to GAPDH. KUNA115 treatment caused a loss of ERK and JNK stimulation over baseline by DAMGO, and a loss of STAT3 protein expression.

Article Snippet: We used the following antibodies for our Western analysis: phospho-Akt (#50-191-224, Fisher Scientific); total-Akt (#50-190-279, Fisher Scientific); phospho-ERK (#50-191-932, Fisher Scientific); total-ERK (#50-191-129, Fisher Scientific); phospho-JNK (#9255, Cell Signaling); total-JNK (#9252, Cell Signaling); Hsp70 (#4872, Cell Signaling); STAT3 (#9139, Cell Signaling); GAPDH (#PIMA515738, Fisher Scientific); Hsp90α (#MA110892, Fisher Scientific); and mCherry (#NBP196752SS, Fisher Scientific).

Techniques: Transduction, Injection, Western Blot, Expressing

Journal: Pharmaceutical Biology

Article Title: Ajudecumin A from Ajuga ovalifolia var. calantha exhibits anti-inflammatory activity in lipopolysaccharide-activated RAW264.7 murine macrophages and animal models of acute inflammation

doi: 10.1080/13880209.2018.1543331

Figure Lengend Snippet: Ajudecumin A repressed the phosphorylation of ERK and p38 MAPK in LPS-stimulated RAW264.7 cells. RAW 264.7 cells were pre-incubated with indicated concentrations of Ajudecumin A for 4 h and then stimulated with 0.5 μg/mL of LPS for another 30 min. An equal amount of protein sample was used to measure the phosphorylation of ERK (A), p38 MAPK (B), and JNK (C) using specific antibodies. The level non-phosphorylated MAPKs protein was used as the internal control. Results from representative experiments are shown, and the quantitative results are depicted. All data are represented as mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. LPS control.

Article Snippet: Phospho- and total-ERK, phosphor- and total-p38 MAPK, phosphor- and total-JNK, phosphor- and total-IκBα antibodies were purchased from Signalway Antibody (Baltimore, USA).

Techniques: Phospho-proteomics, Incubation, Control