Journal: PLoS ONE

Article Title: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines

doi: 10.1371/journal.pone.0188121

Figure Lengend Snippet: (A) Huh7 cells were seeded into a 384-well plate with 1000, 2000, 3000, 4000, and 5000 cells per well. Cell morphology and confluency were observed and an image was captured 72 hours later using a phase contrast microscopy. The arrow indicates the optimized seeding condition for siRNA screening assay. (B) Huh7 cells were reverse transfected with a siNTC or si Caspase 3 for 24 hours before infected with DENV at MOI of 10 for 48 hpi. Conditions were replicated in 15 wells within the same plate. Caspase 3 activity was measured and plotted across the 15 wells.

Article Snippet: Human ON-TARGETplus ® siRNA Library—Apoptosis—SMART pool (Catalogue #G-103900-E2-01) was purchased from GE Dharmacon (Lafayette, CO, USA).

Techniques: Microscopy, Screening Assay, Transfection, Infection, Activity Assay

Journal: PLoS ONE

Article Title: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines

doi: 10.1371/journal.pone.0188121

Figure Lengend Snippet: List of top 20 genes identified from siRNA library screening.

Article Snippet: Human ON-TARGETplus ® siRNA Library—Apoptosis—SMART pool (Catalogue #G-103900-E2-01) was purchased from GE Dharmacon (Lafayette, CO, USA).

Techniques: Activity Assay, Histone Deacetylase Assay

Journal: PLoS ONE

Article Title: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines

doi: 10.1371/journal.pone.0188121

Figure Lengend Snippet: Huh7 cells were reverse transfected with siRNA targeted TNFSF12 or SPHK2 genes or the non-targeting control siRNA for 24 hours before infected with DENV for 48 hours. The mRNA expression was then analyzed using real-time RT-PCR. The mRNA expression of TNFSF12 (A) and SPHK2 (B) represented as fold times compared to parental control. The results are expressed as the average of three independent experiments ± SD. The asterisks indicate statistically significant differences between groups (p < 0.05) (Student’s t test).

Article Snippet: Human ON-TARGETplus ® siRNA Library—Apoptosis—SMART pool (Catalogue #G-103900-E2-01) was purchased from GE Dharmacon (Lafayette, CO, USA).

Techniques: Transfection, Control, Infection, Expressing, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines

doi: 10.1371/journal.pone.0188121

Figure Lengend Snippet: Huh7 cells were reverse transfected with siRNA directed against TNFSF12 or SPHK2 genes for 24 hours before being infected with DENV for 48 hours. Apoptotic cells were determined by Annexin V/PI staining and flow cytometry analysis. (A) Dot plot indicates cell death; and, (B) Apoptotic cells (Annexin V+/PI-) were plotted and compared with percentage of cell population. The results are expressed as the average of three independent experiments ± SD. The asterisks indicate statistically significant differences between groups (p < 0.05) (Student’s t test).

Article Snippet: Human ON-TARGETplus ® siRNA Library—Apoptosis—SMART pool (Catalogue #G-103900-E2-01) was purchased from GE Dharmacon (Lafayette, CO, USA).

Techniques: Transfection, Infection, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: RNAi screen reveals a role of SPHK2 in dengue virus–mediated apoptosis in hepatic cell lines

doi: 10.1371/journal.pone.0188121

Figure Lengend Snippet: Huh7 cells were reverse transfected with siRNA targeted SPHK2 gene for 24 hours before being infected with DENV for 0, 12, 24, 36 and 48 hours. (A) Virus production in supernatant was quantified by FFU assay. (B) DENV E protein expression was determined at 48 hpi using western blot analysis. The results are expressed as the average of three independent experiments ± SD. The asterisks indicate statistically significant differences between groups (p < 0.05) (Student’s t test).

Article Snippet: Human ON-TARGETplus ® siRNA Library—Apoptosis—SMART pool (Catalogue #G-103900-E2-01) was purchased from GE Dharmacon (Lafayette, CO, USA).

Techniques: Transfection, Infection, Virus, Expressing, Western Blot