Journal: Viruses

Article Title: Recombinant Pseudorabies Virus with TK/gE Gene Deletion and Flt3L Co-Expression Enhances the Innate and Adaptive Immune Response via Activating Dendritic Cells

doi: 10.3390/v13040691

Figure Lengend Snippet: Characterization of the recombinant HNX-TK − /gE − -Flt3L. ( A ) Detection of the transcription of the Flt3L gene in HNX-TK − /gE − -Flt3L virus- and HNX-infected cells through RT-qPCR (**** < 0.001). ( B ) Detection of the Flt3L protein and viral gB protein expression through an immunofluorescence assay (IFA). Green: gB-positive cells; red: Flt3L-positive cells; blue: DAPI-stained PK-15 cell nucleus. DAPI, 4’,6-diamidino-2-phenylindole. ( C ) Western blot analysis of β-actin, gD, and Flt3L of the PK-15 cells infected with HNX-TK − /gE − -Flt3L, HNX, or DMEM as a control. ( D ) One step growth curves of HNX-TK − /gE − -Flt3L and HNX in PK-15 cells.

Article Snippet: The blots were blocked by skim milk (BD, Bioscience, Inc., Saint Louis, MO, USA), and this was followed by incubation with rabbit anti-Flt3L polyclonal antibody (Bioss, Beijing, China), mouse anti-gD monoclonal antibody (Keqian Ltd., Wuhan, China), and mouse anti-β-actin monoclonal antibody (Bioss).

Techniques: Recombinant, Infection, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, 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 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