Activation of the Cellular Unfolded Protein Response by Recombinant Adeno-Associated Virus Vectors
Figure 3
Self-complementary AAV2 infection activates PERK1 and IRE1α pathway and its downstream targets.
A. Total RNA from HeLa cells mock-infected or infected with of 5,000 vgs/cell of scAAV2-CB-EGFP vectors was used to profile the expression of downstream targets of IRE1α and PERK target genes such as ATF4 or CHOP by real-time PCR analysis at 2, 6, 12, 24 and 48 hours post infection. *p<0.05 Vs mock infected cells. B. Qualitative reverse-transcription PCR amplification of XBP1 (283 bp) and spliced variant sXBP1 (257 bp) at various time points, 2 h (lane 1), 6 h (lane 2), Molecular weight ladder (lane 3), 12 h (lane 4), 24 h (lane 5) and 48 h (lane 6) analyzed. Dithiothreitol (DTT, lane 7) was used as a positive control of UPR activation.