For cell culture-based qRT-PCR experiments, total RNA was collected
using Monarch Total RNA Miniprep Kits (NEB). For the mouse lung IAV NP qRT-PCR,
lungs were collected at 4 dpi (with 1200 PFU virus) and homogenized directly in
Trizol, before using chloroform and isopropyl alcohol to complete the RNA
extraction. For the confirmation of siRNA knockdown, measurement of ISG RNA
levels, the comparison of relative MSH6 RNA levels in amiRNA-transfected cells,
and the quantification of mouse lung IAV NP levels, samples were analyzed using
the EXPRESS One-Step Superscript Universal qRT-PCR Kit (Thermo) and
gene-specific TaqMan Expression Assay Probes (Thermo) (Supplementary Table 6 ). For the MMR
gene panel, the qRT-PCR was performed in two separate steps. First,
single-stranded cDNA was synthesized using the High-Capacity cDNA Reverse
Transcription Kit with RNase Inhibitor (Thermo). Second, cDNA was loaded into
Custom TaqMan Array 96-well Plates (Thermo) we designed to include all major
genes directly involved in the DNA MMR pathway as well as two housekeeping gene
controls for normalization.
using Monarch Total RNA Miniprep Kits (NEB). For the mouse lung IAV NP qRT-PCR,
lungs were collected at 4 dpi (with 1200 PFU virus) and homogenized directly in
Trizol, before using chloroform and isopropyl alcohol to complete the RNA
extraction. For the confirmation of siRNA knockdown, measurement of ISG RNA
levels, the comparison of relative MSH6 RNA levels in amiRNA-transfected cells,
and the quantification of mouse lung IAV NP levels, samples were analyzed using
the EXPRESS One-Step Superscript Universal qRT-PCR Kit (Thermo) and
gene-specific TaqMan Expression Assay Probes (Thermo) (
gene panel, the qRT-PCR was performed in two separate steps. First,
single-stranded cDNA was synthesized using the High-Capacity cDNA Reverse
Transcription Kit with RNase Inhibitor (Thermo). Second, cDNA was loaded into
Custom TaqMan Array 96-well Plates (Thermo) we designed to include all major
genes directly involved in the DNA MMR pathway as well as two housekeeping gene
controls for normalization.