Figure 2

TRIM25 affects the replication of SADS-CoV. A–D Overexpression of TRIM25 inhibits SADS-CoV replication. IPI-2I cells were transfected with HA-tagged TRIM25 (0, 1, 2, or 4 μg) for 24 h and then infected with SADS-CoV at an MOI of 0.1 for 24 h. A The protein expression of TRIM25 and SADS-CoV N was detected by Western blotting. The densities of SADS-CoV N bands relative to GAPDH were calculated. The value of HA-TRIM25 (0 μg) group was normalized to one. B Overexpression of TRIM25 was verified by qRT-PCR. C The mRNA levels of SADS-CoV N were determined by qRT-PCR. D The SADS-CoV TCID₅₀ in the supernatants was titrated on Vero E6 cells. E-J Knockdown of TRIM25 enhanced SADS-CoV replication. E and F IPI-2I cells were transfected with siTRIM25-1, siTRIM25-2, siTRIM25-3, or siNC (negative control) at 50 nM for 36 h and subsequently infected with SADS-CoV at an MOI of 0.1 for 24 h. The mRNA levels of TRIM25 (E) and SADS-CoV N (F) in TRIM25 knockdown cells were determined by qRT-PCR. G-J IPI-2I cells were transfected with siNC or siTRIM25-1 (50 nM or 100 nM) for 36 h and subsequently infected with SADS-CoV at an MOI of 0.1 for 24 h. G The protein expression of TRIM25 and SADS-CoV N was detected by Western blotting. The densities of TRIM25 and SADS-CoV N bands relative to GAPDH were calculated. The value of negative control cells group was normalized to one. H The mRNA levels of TRIM25 were verified by qRT-PCR. I The mRNA levels of SADS-CoV N were determined by qRT-PCR. J The SADS-CoV TCID₅₀ in the supernatants was titrated on Vero E6 cells. P values were calculated using two-tailed unpaired Student’s t-test.