Journal: Cancer cell

Article Title: Proteome instability is a therapeutic vulnerability in mismatch repair deficient cancer

doi: 10.1016/j.ccell.2020.01.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ERK(1/2) (137F5) , Cell Signaling , Cat# 4695S; RRID: AB 390779.

Techniques: Recombinant, Plasmid Preparation, Expressing, DNA Extraction, Microarray, Sequencing, Mutagenesis, Protein Binding, Synthesized, Functional Assay, shRNA, Software

Journal: JCI Insight

Article Title: Hypochondroplasia gain-of-function mutation in FGFR3 causes defective bone mineralization in mice

doi: 10.1172/jci.insight.168796

Figure Lengend Snippet: ( A ) H&E staining of femur distal growth plate on P14 and P21 showing SOC delay and reduced hypertrophic cartilage area. HZ, hypertrophic zone. Scale bars: 200 μm and 50 μm (zoomed-in images). ( B ) Left: Visualization of SOC of the distal femur. Scale bar: 1000 μm. Right: Graphical representation of the SOC/epiphysis volume ratio in Fgfr3 +/+ ( n = 5) and Fgfr3 Asn534Lys/+ ( n = 5) male and female mice. ( C ) Left: Collagen type X (Col X) immunostaining. Scale bar: 50 μm. Right: Graphical representation of mean cell area in Col X + area in Fgfr3 +/+ ( n = 7) and Fgfr3 Asn534Lys/+ ( n = 6) male and female mice. ( D ) Ki67 + cells/total cells (DAPI + ) on P14 in Fgfr3 +/+ ( n = 6) and Fgfr3 Asn534Lys/+ ( n = 7) male and female mice, BrdU + cells/DAPI + cells on P14 in Fgfr3 +/+ ( n = 7) and Fgfr3 Asn534Lys/+ ( n = 7), and BrdU + cells/DAPI + cells on P21 in Fgfr3 +/+ ( n = 5) and Fgfr3 Asn534Lys/+ ( n = 5) male and female mice. ( E ) Left: p-Erk1/2 immunostaining on P14 growth plate. Scale bar: 50 μm. Graphical representation of relative intensity of p-Erk1/2 on Fgfr3 +/+ ( n = 5) and Fgfr3 Asn534Lys/+ ( n = 7) male and female growth plates. Representative Western blots of p-Erk1/2 (left) and Erk1/2 in primary chondrocytes with FGF2 stimulation over time (0, 5, 30, 60, 120 minutes), from 4 independent Western blots with ( n = 5 mice per group). Graphical representation of p-Erk1/2/Erk1/2 ratio over time. NS, not significant; * P < 0.05 by Mann-Whitney test ( B – D and left graph in E ) or 2-way ANOVA with Šidák’s multiple-comparison test (right graph in E [p-Erk1/2/Erk1/2 ratio over time]).

Article Snippet: Blots were probed with primary antibodies rabbit anti–p-Erk1/2 (Cell Signaling Technology, 4370; 1:2,000), mouse anti–total Erk1/2 (Cell Signaling Technology, 4696; 1:1,000), and mouse anti-actin (Millipore, MAB1501; 1:5,000).

Techniques: Staining, Immunostaining, Western Blot, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Regulation of C6 glioma cell migration by thymol

doi: 10.3892/ol.2016.4237

Figure Lengend Snippet: Western blot analysis of PKCα and ERK1/2 phosphorylation. (A) C6 glioma cells were cultured in serum-free medium for 24 h and then in the presence of thymol (3–30 µM) for 1 h, followed by exposure to 10% FBS for 24 h. Cell lysates were separated by 12% polyacrylamide gel electrophoresis and the specific protein bands were labeled using antibodies. The band intensities were measured and reflect the effect of thymol on the phosphorylation of (B) PKCα and (C) ERK1/2. These values were normalized against total PKCα (T-PKCα), total ERK1/2 (T-ERK1/2) and GAPDH. Data are expressed as the mean ± standard error (n=3). Cell response to FBS is expressed as 100%. *P<0.05 vs. FBS-stimulated cells in the absence of thymol. PKCα, protein kinase Cα; ERK1/2, extracellular signal-regulated kinases 1/2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; FBS, fetal bovine serum.

Article Snippet: Other antibodies, such as rabbit polyclonal anti-rat anti-ERK1/2 (catalog no., sc-94), mouse monoclonal anti-rat anti-P-ERK1/2 (catalog no., sc-7383), rabbit polyclonal anti-rat anti-MMP2 (catalog no., sc-10736) and goat polyclonal anti-rat anti-MMP9 (catalog no., sc-6840), were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Western Blot, Cell Culture, Polyacrylamide Gel Electrophoresis, Labeling