Mitochondrial protein, TBRG4, modulates KSHV and EBV reactivation from latency
Fig 1
Knockdown of TBRG4 increases KSHV lytic reactivation.
iSLK.219 cells were transfected with control non-specific (NS) siRNA or TBRG4 siRNA for 48 h and then treated with doxycycline (Dox; 0.2 μg/mL). (A) KSHV genome copy numbers in the supernatant were measured by real-time PCR at the indicated time. (B-F) RNA was extracted from the cells, and the knockdown efficiency of TBRG4 and the mRNA expression levels of the KSHV ORF57, ORF39, K8.1, and ORF52 genes were measured by real-time PCR. Target mRNA expression was normalized to β-actin mRNA and presented as fold induction. (G) Cell lysates were collected at the indicated time points, and western blots were performed with the indicated antibodies. (H) RNA was extracted from duplicate samples at the indicated time points, and KSHV viral transcript expression was measured by a KSHV real-time PCR-based array targeting 82 KSHV viral genes. Target viral mRNA expression levels were normalized to the mRNA level of cellular housekeeping genes to generate delta CT as a measure of relative expression. A heatmap for the KSHV viral array using the delta CT value was shown. Higher transcript levels are indicated by red, and lower levels by blue. Viral genes are listed on the vertical axis and samples on the horizontal. R1 and R2 indicate data from duplicate samples. The data shown are representative of three independent experiments. Data are presented as mean ± SD. *, P<0.05 by student’s t-test.