Journal:

Article Title: Simultaneous gene expression analysis of steady-state and actively translated mRNA populations from osteosarcoma MG-63 cells in response to IL-1? via an open expression analysis platform

doi: 10.1093/nar/gkg702

Figure Lengend Snippet: (A) RT–QPCR analysis of PP-2A expression in MG-63 cells. The average CT values for PP-2A were first normalized against the housekeeping gene GAPDH and converted to a percentage for relative expression (n = 3). (B) Western immunoblot analysis of PP-2A in MG-63 cells. Cytosolic extracts from MG-63 cells (lane 1) and MG-63 cells treated with IL-1α (lane 2) were prepared. PP-2A protein was detected using an anti-PP-2A mouse monoclonal antibody. Filtered membranes were then reprobed with an anti-β-actin monoclonal antibody to control for loading and integrity of protein.

Article Snippet: Proteins were probed with rabbit anti-CAML polyclonal antibody (1:4000 dilution), mouse anti-β-actin monoclonal antibody (1:5000 dilution) and mouse anti-PP-2A antibody (1:1000 dilution), followed by incubation with a horseradish peroxidase-conjugated secondary antibody (1:1000 dilution) (Bio-Rad, CA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal:

Article Title: Simultaneous gene expression analysis of steady-state and actively translated mRNA populations from osteosarcoma MG-63 cells in response to IL-1? via an open expression analysis platform

doi: 10.1093/nar/gkg702

Figure Lengend Snippet: (A) RT–QPCR analysis of CAML expression in MG-63 cells. CT values for CAML were first normalized against housekeeping gene GAPDH and converted to a percentage for relative expression. (B) Western immunoblot analysis of CAML in MG-63 cells. Cytosolic extracts from MG-63 cells (lane 2) and MG-63 cells treated with IL-1α (lane 1) were prepared. CAML protein was detected by immunoblot analysis using an anti-CAML polyclonal antibody. Filtered membranes were then reprobed with an anti-β-actin monoclonal antibody to control for loading and integrity of protein.

Article Snippet: Proteins were probed with rabbit anti-CAML polyclonal antibody (1:4000 dilution), mouse anti-β-actin monoclonal antibody (1:5000 dilution) and mouse anti-PP-2A antibody (1:1000 dilution), followed by incubation with a horseradish peroxidase-conjugated secondary antibody (1:1000 dilution) (Bio-Rad, CA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Disruption of the FasL/Fas axis protects against inflammation-derived tumorigenesis in chronic liver disease

doi: 10.1038/s41419-019-1391-x

Figure Lengend Snippet: a Representative TUNEL staining of 8-week-old NEMO f/f , NEMO Δhepa , Fas lpr , and NEMO Δhepa /Fas lpr livers. b TUNEL-positive cells were quantified and graphed. c Proliferation was determination by Ki-67-positive cells immunofluorescence. d Ki-67-positive cells were quantified and graphed. e Expression of PCNA, cleaved Caspase-3 (CC3), and RIPK1 was analysed by immunoblotting of whole liver extracts. GAPDH served as a loading control. Results are expressed as mean ± SEM ( n = 8 livers, p < 0.001–0.05)

Article Snippet: Isolated protein samples were probed with antibodies against RIPK1 (#5389) (Pro-Sci, Poway, CA, USA), Cleaved Caspase-3 (Asp175) (Cell Signaling Technology, Massachusetts, USA), PCNA (clone PC10) (Dianova GmbH, Hamburg, Germany), and GAPDH (MCA4739; 1:5000; AbD SeroTec, Düsseldorf, Germany).

Techniques: TUNEL Assay, Staining, Immunofluorescence, Expressing, Western Blot, Control