mouse mab sc 1668  (Santa Cruz Biotechnology)

Journal: PLoS Pathogens

Article Title: Consecutive Inhibition of ISG15 Expression and ISGylation by Cytomegalovirus Regulators

doi: 10.1371/journal.ppat.1005850

Figure Lengend Snippet: (A) Normal HF cells or cells expressing control (shC) or UBP43-specific shRNAs (shUBP43-1 and -2) were treated with IFNα (2,000 U/ml) for 24 h and the UBP43 levels were analyzed by immunoblotting using anti-UBP43 antibody. The tubulin levels were shown as a loading control. Arrowheads indicate two UBP43 bands with different molecular weights and an open circle indicates non-specific bands. (B) Control and UBP43 knockdown HF cells were infected or not with UV-HCMV at an MOI of 3. At 24 h after infection, cell lysates were immunoblotted with antibodies for ISG15 and β-actin. (C) Control and UBP43-knockdown HF cells were infected with HCMV at an MOI of 3. At 5 days after infection, viral supernatants were collected and the levels of progeny virions were measured by infectious center assays. Scatter plots are shown. (D) Control and UBP43-knockdown cells were infected as in (C). Cell lysates were prepared at indicated time points and immunoblotted with antibodies for ISG15, STAT2, phosphorylated STAT2 (on Tyr 689), and β-actin. (E) Control HF cells or cells expressing Herc5-specific shRNA were mock-infected or infected with UV-HCMV at an MOI of 3. The Herc5 transcript levels were determined at 24 h after infection by qRT-PCR. The β-actin mRNA levels were used for normalization. Values are an average of duplicated assays; error ranges are indicated. (F) Control and Herc5-knockdown HF cells were infected or not with UV-HCMV at an MOI of 3. At 24 h after infection, cell lysates were immunoblotted with antibodies for ISG15 and β-actin. (G) Control and Herc5-knockdown HF cells were infected with HCMV at an MOI of 0.1. At 9 days after infection, virus titers in the culture supernatants were measured by infectious center assays.

Article Snippet: Anti-ISG15 (F-9) and anti-STAT2 mouse MAbs were obtained from Santa Cruz.

Techniques: Expressing, Control, Western Blot, Knockdown, Infection, shRNA, Quantitative RT-PCR, Virus