Journal: Cells

Article Title: Hepatitis C Virus Dysregulates Polyamine and Proline Metabolism and Perturbs the Urea Cycle.

doi: 10.3390/cells13121036

Figure Lengend Snippet: Scheme 1. Scheme of the metabolism of biogenic polyamines and adjacent pathways and their pharmacological inhibitors used in this study. Compounds that target metabolic enzymes are pre- sented in red. ARG—arginase, OTC—ornithine transcarbamoylase, ASS—argininosuccinate synthase, ASL—argininosuccinate lyase, ODC—ornithine decarboxylase, AMD—S-adenosylmethionine decar- boxylase, SRM—spermidine synthase, SMS—spermine synthase, SSAT—spermidine/spermine-N1- acetyltransferase, PAOX—acetylpolyamine oxidase, SMOX—spermine oxidase, PRODH—proline dehy- drogenase, P5C—∆1-pyrrolidine-5-carboxylate, PYCR—P5C reductase, ALDH—aldehyde dehydrogenase.

Article Snippet: Primary murine antibodies to β-actin (ab3280, 1:500), ASS (ab124465, 1:2000), and ODC1 (ab193338, 1:1000), as well as rabbit antibodies to ASL1 (ab97370, 1:2000), were from Abcam (Cambridge, UK); rabbit antibodies to arginase I (9819s, 1:1000) and SSAT (61586, 1:200) were obtained from Cell Signaling Technology (Danvers, MA, USA), and antibodies to SMOX (hpa047117, 1:1000) from Sigma (St. Louis, MO, USA).

Techniques:

Journal: Cells

Article Title: Hepatitis C Virus Dysregulates Polyamine and Proline Metabolism and Perturbs the Urea Cycle.

doi: 10.3390/cells13121036

Figure Lengend Snippet: Figure 1. HCV infection suppresses the expression of polyamine-metabolizing enzymes at the posttranscriptional level. (A–F) Huh7.5 cells were infected with HCV at MOI 0.1. (A) Infection spread was monitored by immunofluorescence staining using anti-NS3 rabbit sera and FITC-labelled antirabbit antibodies; the nuclei were counterstained with DAPI, and signals were visualized by confocal microscopy at 63× magnification. The yellow scale bar corresponds to 25 µm; the red bars correspond to 50 µm. (B–D) Expression of polyamine-metabolizing proteins was assessed 3–10 days post-infection (dpi) by RT-qPCR (B,C) or Western blotting (D). Intracellular SSAT activity was quantified by monitoring the transfer of the 14C-labelled acetyl group from [14C]-AcCoA to the spermine molecule. (F) Polyamine levels were measured by HPLC. Graphs represent means ± SEM of four (B) or three (C,E,F) independent experiments. * p < 0.05, ** p < 0.01, # p < 0.1 compared to mock-infected cells by ANOVA with Dunnett’s post hoc test (B,C,F) or paired t-test (E).

Article Snippet: Primary murine antibodies to β-actin (ab3280, 1:500), ASS (ab124465, 1:2000), and ODC1 (ab193338, 1:1000), as well as rabbit antibodies to ASL1 (ab97370, 1:2000), were from Abcam (Cambridge, UK); rabbit antibodies to arginase I (9819s, 1:1000) and SSAT (61586, 1:200) were obtained from Cell Signaling Technology (Danvers, MA, USA), and antibodies to SMOX (hpa047117, 1:1000) from Sigma (St. Louis, MO, USA).

Techniques: Infection, Expressing, Immunofluorescence, Staining, Confocal Microscopy, Quantitative RT-PCR, Western Blot, Activity Assay