RNA Sequencing:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
Knockdown:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
Cell Culture:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
Transfection:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
Control:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
Modification:Article Title: Identification of host lncRNAs that impact Venezuelan equine encephalitis virus TC-83 replication.
Article Snippet: .. RNA sequencing of Snhg15 knockdown cells Three biological replicates of primary mouse astrocytes (Lonza, M-ASM-330) at passage five were cultured into six-well plates and maintained in astrocyte growth medium at 37°C and 5% CO2 for 24 h. At 24-h post-seeding, the cells were transfected with either siRNA targeting Snhg15 or a pool of four non-targeting siRNAs (Control) using TransIT-X2 transfection reagent (Mirus, MIR6004) according to the manufacturer’s protocol with modification of using half of the suggested transfection reagent. .. The transfected cells were incubated at 37°C and 5% CO2 for 24 h; 24 h after transfection, the cells were incubated with VEEV TC-83 (MOI 5) for 1 h with rocking at 30 min. After 1 h, the viral inoculum was replaced with cell culture medium, and cells were incubated at 37°C and 5% CO2 for another 24 h. At 24 h post-infection, the cells were lysed using Tri-reagent (Zymo Research, R2050-1-200), and total RNA was extracted from the cell lysates using Direct-Zol RNA Microprep Kit (Zymo Research, R2060) according to the manufacturer’s protocol.
|