FIG 6.

RNA transcription, protein translation, interferon responses, and NSP4-VP6 interactions are unaffected by N-glycosylations. (A) Viral RNA kinetics in cells infected with rsSA11 or rsSA11-Y9P-N18A. HT29 cells were infected with rsSA11 or rsSA11-Y9P-N18A at an MOI of 5. Cell lysates were collected at 0, 4, 8, 12, and 24 hpi. RNA was extracted and subjected to qRT-PCR. The data are expressed as means ± the SD of triplicate samples. (B) Serial Western blots. HT29 cells were infected with rsSA11 or rsSA11-Y9P-N18A at an MOI of 5, and lysates were collected at the designated time points. The molecular weight (kDa) is shown on the right. (C) IFN responses. HT29 cells were infected with rsSA11 or rsSA11-Y9P-N18A at an MOI of 5. At 8 hpi, cell lysates were collected and subjected to qRT-PCR. Relative mRNA levels (compared to mock samples) are shown (mock sample was set as 1). The data are expressed as means ± the SD of triplicate samples. Statistical significance was determined using a t test, with P < 0.05 considered significant (*, P < 0.05). (D) NSP4-VP6 interaction. HT29 cells were infected with rsSA11 or rsSA11-Y9P-N18A at an MOI of 5. At 18 hpi, cell lysates were collected and subjected to coimmunoprecipitation with an anti-NSP4 antibody. The molecular weight is shown on the right (IP, immunoprecipitation; WCL, whole-cell lysate).