Parvovirus B19 NS1 protein induces cell cycle arrest at G2 phase

We construct two stable UT7/Epo-S1 cell lines which could be inducible expressing B19 NS1 and NS1 TAD2 domain mutation proteins. After treated with or without doxycycline induction, we extract the total RNA for RNA-seq analysis. UT7/Epo-S1 cells were transduced by lentiviruses inducible expressing NS1 or NS1mTAD2. Two days post-transduction, puromycin was added at a concentration of 0.5 μg/ml, then cells were cultured in presence of puromycin for 3 to 4 passages to select the lentivirus transduced cells. UT7/Epo-S1 cells stably expressing NS1 (NS1-S1) and NS1mTAD2 (NS1mTAD2-S1) were cultured under similar conditions as normal S1 cells except for the addition of 5 mg/ml doxycycline (Dox) for inducing the expression of NS1 and NS1mTAD2 proteins, respectively. Total RNA of the two cell lines treated with or without Dox were extracted and for further RNA-seq analysis.