Journal: Virology

Article Title: The interplay between viperin antiviral activity, lipid droplets and Junín mammarenavirus multiplication.

doi: 10.1016/j.virol.2017.10.012

Figure Lengend Snippet: Fig. 6. LD characterization in JUNV infected HepG2 cells. (A) HepG2 cells were infected at MOI of 0.1 PFU/cell and expression of RIG-I and viperin genes was determined at 24 h and 48 h p.i. (B) Control uninfected and JUNV infected HepG2 cultures were fixed and probed for LDs detection and viral G1 viral antigen. Representative images of cultures probed with Plin2 (green channel) and G1 (red channel). (C) LDs quantification of JUNV (+) cells over negative cells. Brightfield and merge of all channels are also shown. Magnification 600X. ns: non significative. (*) significative values.

Article Snippet: A549 (human lung adenocarcinoma; ATCC CL-185), HepG2 (human hepatoma; ATCC CRL-10741) and Vero (African green monkey kidney; ATCC CCL 81) cell lines were grown in Eagle's minimum essential medium (MEM) (GIBCO) supplemented with 10%, 20% and 5% of fetal bovine serum respectively and 50 μg/ml gentamycin.

Techniques: Infection, Expressing, Control

Journal: Virology

Article Title: The interplay between viperin antiviral activity, lipid droplets and Junín mammarenavirus multiplication.

doi: 10.1016/j.virol.2017.10.012

Figure Lengend Snippet: Fig. 7. Effect of C75 on JUNV multiplication. (A) HepG2 cells were incubated for 48 h with different concentrations of C75 and then cell viability was determined by MTT and crystal violet assays. (B) Untreated (CC) and 23 μM C75 treated HepG2 cells were fixed and probed with BODIPY. LD abundance was quantified as explained in Materials and methods. Magnification: 600X. (C) JUNV suspensions were incubated with increasing concentrations of C75 and remaining JUNV infectivity was determined by plaque assay. (D) JUNV infected cultures were incubated in presence or absence of C75 and virus yields were determined at 48 h p.i.

Article Snippet: A549 (human lung adenocarcinoma; ATCC CL-185), HepG2 (human hepatoma; ATCC CRL-10741) and Vero (African green monkey kidney; ATCC CCL 81) cell lines were grown in Eagle's minimum essential medium (MEM) (GIBCO) supplemented with 10%, 20% and 5% of fetal bovine serum respectively and 50 μg/ml gentamycin.

Techniques: Incubation, Infection, Plaque Assay, Virus

Journal: Virology

Article Title: The interplay between viperin antiviral activity, lipid droplets and Junín mammarenavirus multiplication.

doi: 10.1016/j.virol.2017.10.012

Figure Lengend Snippet: Fig. 8. Subcellular localization of viperin and viral proteins. HepG2 cultures were infected with JUNV and at 48 h p.i. cells were fixed, probed with BODIPY and stained with mouse monoclonal antibodies to reveal (A) JUNV G1 protein or (B) JUNV N protein. Magnification: 600×.

Article Snippet: A549 (human lung adenocarcinoma; ATCC CL-185), HepG2 (human hepatoma; ATCC CRL-10741) and Vero (African green monkey kidney; ATCC CCL 81) cell lines were grown in Eagle's minimum essential medium (MEM) (GIBCO) supplemented with 10%, 20% and 5% of fetal bovine serum respectively and 50 μg/ml gentamycin.

Techniques: Infection, Staining, Bioprocessing

Journal: Virology

Article Title: The interplay between viperin antiviral activity, lipid droplets and Junín mammarenavirus multiplication.

doi: 10.1016/j.virol.2017.10.012

Figure Lengend Snippet: Fig. 9. Interaction of viperin and JUNV N proteins. (A) Infected HepG2 cells were stained with mouse monoclonal anti-viperin antibodies and rabbit polyclonal anti-JUNV antibodies, and further secondary antibodies incubation as detailed in Materials and methods. (B) A549 cells cultures were infected with JUNV at MOI of 0.1 PFU/cell. At 24 or 48 h p.i. monolayers were processed for immunoprecipitation anti-N (NA05AG12) antibodies. Immunocomplexes were subjected to WB analysis to detect viperin presence with polyclonal antibodies. As a control non infected cells (CC) collected with lysis buffer containing and processed as infected samples. (C) Co-localization of viperin- JUNV N protein. Representative image of co- transfected A549 cells stained with anti-viperin (green channel) and anti-N (red channel) antibodies. Merge panel shows co-localization overlap spots with a Pearson coefficient = 0.65. (D) HEK293T cells were transfected with plasmids encoding FLAG-tagged viperin and HA-tagged Junin N protein in different combinations as indicated (+). At 24 h p.t., cell extracts were subjected to anti-HA or anti-Flag immunoprecipitation. Whole cell lysate (input) and immunoprecipitated proteins were analyzed by immunobloting using antibodies as indicated. Tubulin served as a loading control. Representative images are shown from three independent experiments.

Article Snippet: A549 (human lung adenocarcinoma; ATCC CL-185), HepG2 (human hepatoma; ATCC CRL-10741) and Vero (African green monkey kidney; ATCC CCL 81) cell lines were grown in Eagle's minimum essential medium (MEM) (GIBCO) supplemented with 10%, 20% and 5% of fetal bovine serum respectively and 50 μg/ml gentamycin.

Techniques: Infection, Staining, Incubation, Immunoprecipitation, Control, Lysis, Transfection, Western Blot

Journal: Nanoscale Research Letters

Article Title: Assessment of Gold Nanoparticles-Inhibited Cytochrome P450 3A4 Activity and Molecular Mechanisms Underlying Its Cellular Toxicity in Human Hepatocellular Carcinoma Cell Line C3A

doi: 10.1186/s11671-018-2684-1

Figure Lengend Snippet: C3A viability and LC 50 values of the 40 and 80 nm a AuNP and b PC AuNP. Data represent mean ± S.D. ( n = 3). PC human plasma protein corona, ND not determined, BPEI branched polyethylenimine, LA lipoic acid, PEG polyethylene glycol, LC 50 median lethal concentration

Article Snippet: Human hepatocellular carcinoma C3A cells (ATCC ® CRL-10741TM) were purchased from ATCC ® (Manassas, VA), cultured in complete EMEM (ATCC ® , Manassas, VA) supplemented with 10% FBS, and expanded to approximately 80% confluence in T75 flask with medium changes every 4 days.

Techniques: Clinical Proteomics, Concentration Assay

Journal: Nanoscale Research Letters

Article Title: Assessment of Gold Nanoparticles-Inhibited Cytochrome P450 3A4 Activity and Molecular Mechanisms Underlying Its Cellular Toxicity in Human Hepatocellular Carcinoma Cell Line C3A

doi: 10.1186/s11671-018-2684-1

Figure Lengend Snippet: Time-dependent cellular uptake of the 40 nm a BPEI-AuNP, b LA-AuNP, and c PEG-AuNP, and the 80 nm d BPEI-AuNP, e LA-AuNP, and f PEG-AuNP in the absence and presence of PC in C3A cells up to 24 h. Data represent mean ± S.D. ( n = 3). Letters were significantly different according to Tukey’s HSD test. BPEI branched polyethylenimine, LA lipoic acid, PEG polyethylene glycol, PC human plasma protein corona, MSD minimum significant difference. * p < 0.05; ** p < 0.005; *** p < 0.0001

Article Snippet: Human hepatocellular carcinoma C3A cells (ATCC ® CRL-10741TM) were purchased from ATCC ® (Manassas, VA), cultured in complete EMEM (ATCC ® , Manassas, VA) supplemented with 10% FBS, and expanded to approximately 80% confluence in T75 flask with medium changes every 4 days.

Techniques: Clinical Proteomics

Journal: Nanoscale Research Letters

Article Title: Assessment of Gold Nanoparticles-Inhibited Cytochrome P450 3A4 Activity and Molecular Mechanisms Underlying Its Cellular Toxicity in Human Hepatocellular Carcinoma Cell Line C3A

doi: 10.1186/s11671-018-2684-1

Figure Lengend Snippet: Time- and concentration-dependent ROS/RNS production in C3A cells exposed to a the 40 nm BPEI-AuNP and b the 40 nm PEG-AuNP up to 24 h. Data represent mean ± S.D. ( n = 3). Letters were significantly different according to Tukey’s HSD test. BPEI branched polyethylenimine, LA lipoic acid, PEG polyethylene glycol, CTRL control, MSD a minimum significant difference, PCN pyocyanin (ROS inducer). ** p < 0.005; *** p < 0.0001

Article Snippet: Human hepatocellular carcinoma C3A cells (ATCC ® CRL-10741TM) were purchased from ATCC ® (Manassas, VA), cultured in complete EMEM (ATCC ® , Manassas, VA) supplemented with 10% FBS, and expanded to approximately 80% confluence in T75 flask with medium changes every 4 days.

Techniques: Concentration Assay, Control

Journal: Nanoscale Research Letters

Article Title: Assessment of Gold Nanoparticles-Inhibited Cytochrome P450 3A4 Activity and Molecular Mechanisms Underlying Its Cellular Toxicity in Human Hepatocellular Carcinoma Cell Line C3A

doi: 10.1186/s11671-018-2684-1

Figure Lengend Snippet: An inhibitory effect of AuNP on CYP3A4 activity in C3A cells exposed to the 40 and 80 nm BPEI-, LA-, and PEG-AuNP in the absence and presence of PC for 24 h. Values represent mean ± S.D. ( n = 3). BPEI branched polyethylenimine, LA lipoic acid, PEG polyethylene glycol, PC human plasma protein corona

Article Snippet: Human hepatocellular carcinoma C3A cells (ATCC ® CRL-10741TM) were purchased from ATCC ® (Manassas, VA), cultured in complete EMEM (ATCC ® , Manassas, VA) supplemented with 10% FBS, and expanded to approximately 80% confluence in T75 flask with medium changes every 4 days.

Techniques: Activity Assay, Clinical Proteomics