Figure 3
From: Functional and molecular dissection of HCMV long non-coding RNAs
HCMV lncRNAs other than RNA5.0 are necessary for latent reservoir maintenance and efficient reactivation in Kasumi-3 cells. (a) Schematic representation of latency and reactivation analysis in Kasumi-3 cells using WT and lncRNA-Mut viruses. (b) The levels of viral DNA at the indicated time points were measured by qPCR of the UL123 region, and normalized against GAPDH. (c, d) The RNA levels of LUNA (c), and UL138 (d) at the indicated time points were quantified using qRT-PCR. (e) WT or lncRNA-Mut HCMV-infected Kasumi-3 cells were treated with DMSO or 20 nM PMA and co-cultured with naïve HFF cells. Frequency of infectious centers was calculated using ELDA. (f) Representative images of HFF cells co-cultured with 1600 PMA-treated Kasumi-3 cells for 14 days. Each image is obtained from each well of HFFs. IE1/2-positive images are presented on the left. Merged images are shown for IE1/2 (green), and DAPI (blue) staining. (b–e) Data represent mean ± SEM. All experiments were performed independently (n = 3), and the statistical significance was calculated by two-tailed unpaired t-test.
