DOI: 10.5061/dryad.pk0p2nh1j
title: Modulation of DNA methyltransferases (DNMTs) in Spodoptera frugiperda (Sf9) cells following AcMNPV infection and its effects on the virus-cell interaction

description: In this study, we examined the gene expression profiles of Sf9 insect
 cells following infection with baculovirus, utilizing reverse
 transcription quantitative PCR (RT-qPCR) to analyze how the host cellular
 machinery responds at the transcriptional level to viral invasion. Our
 findings highlight the intricate regulatory mechanisms in place during
 virus replication. To further understand the relationship between the host
 response and viral propagation, we also quantified the dynamics of
 baculovirus replication within the Sf9 cells using quantitative PCR
 (qPCR). This dual approach provides a comprehensive view of both the
 efficiency of viral replication and the accompanying host transcriptional
 changes.
description: # Modulation of DNA methyltransferases (DNMTs) in Spodoptera frugiperda
 (Sf9) cells following AcMNPV infection and its effects on the virus-cell
 interaction Dataset DOI:
 [10.5061/dryad.pk0p2nh1j](10.5061/dryad.pk0p2nh1j) ## Description of the
 data and file structure **File: data.txt** Description: Includes a text
 file of tables. To analyze gene expression levels in AcMNPV-infected and
 control uninfected Sf9 cells, total RNA was extracted from cells at 4, 8,
 16, 24 and 48 hpi. Also, to determine the effect of 5-AZA on the
 expression of immune-related genes, RNA samples were extracted at 24 and
 48 h post AcMNPV infection. Total RNA was extracted by Tri- Reagent™
 according to the manufacturer's instructions (Molecular Research
 Center Inc.). RNA samples were treated with DNase I and incubated at 37 °C
 for 10 min followed by heat inactivation at 75 °C for 10 min. RNA
 concentrations were measured using an Epoch microplate spectrophotometer
 (BioTek) and integrity was ensured through analysis of RNA on a 1% (w/v)
 agarose gel. The first strand cDNA was synthetized using MMLV-reverse
 transcriptase enzyme (Promega) and an oligo dT primer using 1μg of total
 RNA as template at 42 °C for 60 min followed by heating at 65 °C for 5
 min. qPCR was performed by utilizing the SYBR Green Mix without ROX
 (Ampliqon) with a Mic real-time PCR (BMS) under the following conditions:
 95 °C for 15min, followed by 40 cycles of 95 °C for 10s, 57 °C for 10 s,
 and 72 °C for 20 s, followed by the melting curve (60–95 °C). In all
 experiments, ribosomal protein L27 (*Rpl27*) was used as the reference
 gene and also three biological replicates with three technical replicates
 were analyzed for each experiment.  QPCR data were analyzed using the
 relative expression ratio method [ratio = (*E*target)ΔCPtarget(control –
 sample)/(*E*ref)ΔCPref(control – sample)]. This data quantifies how host
 cell gene expression and viral replication change over time after
 baculovirus infection, and shows how treatment with a demethylating agent
 (Aza-5-azacytidine) alters expression of selected genes. The data shown in
 each table represent normalized gene expression results (three replicates)
 at hours post-infection. The mean and standard deviation for each time
 point are also indicated in the table. Each table corresponds to the
 specified gene (DNMT1, DNMT2, Rel – Relish, Dor – Dorsal, Glo – Gloverin,
 Def – Defensin). The replication level of AcMNPV in Sf9 cells at hours
 post-infection is also indicated. In this table, the normalized virus
 replication results (three replicates) at hours post-infection are shown.
 The mean and standard deviation for each time point are also indicated in
 the table.