FIGURE 1.

Porcine hemagglutinating encephalomyelitis virus (PHEV) infection up-regulates miR-10a-5p expression. (A) MiRNA array of PHEV-infected and uninfected mice. Hierarchical clustering shows distinguishable miRNA expression profiling among samples. (B) N2a cells were infected with PHEV at 2 × 10⁶ TCID₅₀/ml and harvested at 0, 24, 48, and 72 h post infection. The expression of miR-10a-5p was determined by qRT-PCR. U6 was chosen as a housekeeping gene to normalize miR-142a-3p expression. Another irrelative miR-130b-5p is used as a negative control. (C) Cells were treated as described in panel (B), then the total RNA was extracted, and the relative expression of PHEV mRNA was determined by qRT-PCR. Data were normalized to GAPDH. (D) N2a cells treated as described in panel (B), and the cell lysates were collected and subjected to examine the expression of PHEV protein by western blotting. Beta actin expression was verified as loading control. (E) BALB/c mice were infected with PHEV for 3 or 5 days, and then the cerebral cortexes were harvested, and miR-10a-5p level was determined by qRT-PCR. N = 4 mice per group, three independent experiments. (F) The lysis from cerebral cortexes of mice as described in panel (E) was collected, and then the relative expression of PHEV mRNA was determined by qRT-PCR. (G) The cerebral cortical lysis was detected by western blotting, and the PHEV protein was normalized to beta actin. All of data are representative of at least three independent experiments, with the error bars representing the standard deviations (SDs). *p < 0.05, **p < 0.01 vs. normal controls.