Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were pre-treated with varying concentrations of andrographolide, or with vehicle only, or not treated and then infected or mock infected with ( a ) CHIKV E1: 226VT ( b ) CHIKV E1: 226AS and ( c ) E1: 226VT followed by incubation under standard conditions in the presence or absence of the drug or vehicle as appropriate. At 24 h.p.i. the level of ( a ) infection and cell viability were determined by flow cytometry and trypan blue staining or ( b , c ) viral production determined by standard plaque assay. Experiments were undertaken independently in triplicate, with duplicate plaque assay. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Infection, Incubation, Flow Cytometry, Staining, Plaque Assay

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with varying concentrations of andrographolide, or with vehicle only or not treated and then infected with CHIKV E1: 226VT followed by incubation under standard conditions in the presence or absence of the drug or vehicle as appropriate. At 24 h.p.i. cells were fixed and stained to show the nucleus (red) and CHIKV E2 protein (green). Cells were examined under an Olympus FluoView 1000 confocal microscope with 60X magnification. Representative, non-contrast adjusted merged images are shown.

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Staining, Microscopy

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with varying concentrations of andrographolide or with vehicle only or not treated and then infected with CHIKV E1: 226VT or mock infected followed by incubation under standard conditions in the presence or absence of the drug or vehicle as appropriate. At 24 h.p.i. samples were examined by western blotting for the expression of CHIKV structural and non-structural proteins.

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with varying concentrations of andrographolide, or with vehicle only or not treated and then mock infected or infected with ( a ) CHIKV E1: 226VT or ( b ) E1: 226AS followed by incubation under standard conditions in the presence or absence of the drug or vehicle as appropriate. At 24 h.p.i., CHIKV RNA copy number was determined by qRT-PCR. Experiments were undertaken independently in duplicate with duplicate qRT-PCR. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with 100 μM andrographolide or with vehicle only or not treated at the indicated time points before and after mock infection or infection with ( a , b ) CHIKV E1: 226VT or ( c , d ) CHIKV E1: 226AS. At ( a to c ) 24 or ( d ) 36 h.p.i. ( a ) cells were collected to determine the cell viability by trypan blue staining and infection level by flow cytometry or ( b to d ) supernatants were collected to determine virus titer by standard plaque assay. Experiments were undertaken independently in triplicate with duplicate plaque assay. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Staining, Flow Cytometry, Virus, Plaque Assay

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with 100 μM andrographolide or with vehicle only or not treated at the indicated time points before and after mock infection or infection with CHIKV E1: 226VT. At 24 h.p.i. cells were fixed and stained to show the nucleus (red) and CHIKV E2 protein (green). Cells were examined under an Olympus FluoView 1000 confocal microscope with 60X magnification. Representative, non-contrast adjusted merged images are shown.

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Staining, Microscopy

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with 100 μM andrographolide or with vehicle only or not treated at the indicated time points before ( a , c , e ) or after ( b , d , e ) mock infection or infection with CHIKV: E1 226VS ( a , b ) or E1: 226AS ( c to f ) and at a time point of 24 ( a to d ) or 36 ( e , f ) hours after infection the expression of structural (Capsid, E1, pE2 and E2) and non-structural (nsP2, nsP3) CHIKV proteins together with actin as a control was analyzed by western blotting. Experiments were undertaken independently in duplicate.

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Expressing, Control, Western Blot

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were incubated with 100 μM andrographolide or with vehicle only or not treated at the indicated time points before or after mock infection or infection with CHIKV: E1 226VS ( a ) or E1: 226AS ( b , c ). At 24 ( a , b ) or 36 ( c ) hours after infection CHIKV RNA copy number was quantitated by qRT-PCR. Experiments were undertaken in duplicate with duplicate qRT-PCR. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Incubation, Infection, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were directly transfected with CHIKV E1: 226AS ( a , c ) or E1: 226VS ( b , d ) RNA or mock transfected and then treated with varying concentrations of andrographolide or vehicle or not treated. At 24 h.p.i., ( a , b ) cellular expression of E2/pE2 and actin were determined by western blotting and ( c , d ) titer of CHIKV in the supernatants were determined by standard plaque assay. Experiments were undertaken independently in duplicate with duplicate of plaque assay. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Transfection, Expressing, Western Blot, Plaque Assay

Journal: Scientific Reports

Article Title: Activity of andrographolide against chikungunya virus infection

doi: 10.1038/srep14179

Figure Lengend Snippet: HepG2 cells were pre-treated with varying concentrations of andrographolide or with vehicle only or not treated and then infected or mock infected with ( a ) CHIKV E1: 226VT ( b ) CHIKV E1: 226AS and ( c ) E1: 226VT followed by incubation under standard conditions in the presence or absence of the drug or vehicle as appropriate. At 24 h.p.i. ( a ) the level of viral production was determined by standard plaque assay and ( b ) the genome copy number in cells was determined by qRT-PCR. The experiments were undertaken independently in duplicate with duplicate plaque assay or qRT-PCR as appropriate. Bars show mean +/− SD (*; p value < 0.05).

Article Snippet: The human liver cell line HepG2 (ATCC Cat No. HB-8065) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated fetal calf serum (FCS) at 37 °C with 5% CO 2.

Techniques: Infection, Incubation, Plaque Assay, Quantitative RT-PCR