Figure 2
Evaluation of apoptosis and assessment of variety of genes and molecular events associated with intrinsic and extrinsic pathways of apoptosis in Huh7 cells transfected with full-length linear monomers of HBV/D1, D2, D3 and D5. Apoptotic cell death was determined by Annexin-V-FITC and propidium idodide (PI) staining method using flow cytometry. (A) A representative experiment showing percentage of apoptotic cells. In each panel the lower left quadrant shows both Annexin-V and PI negative cells (live cells), lower right quadrant shows only Annexin-V positive cells (early apoptotic cells), upper right quadrant shows both Annexin-V and PI positive cells (Late apoptotic cells), upper left quadrant shows only PI positive cells (necrotic cells). The sum of the percentages of both early and late apoptotic cells in presence of different D-subgenotype was determined and the data was represented in mean ± SD bar (n = 3). Relative expression of (B) caspase-3, (C) Bax, (D) Bcl-2, (E) TNF-R1, (F) FasR, (G) TRAIL-R1, (H) Grp78, (I) ATF4 and (J) CHOP were evaluated by real-time PCR in Huh7 cells transfected with D-subgenotype. The levels of (K) ROS in D-subgenotype transfected Huh7 cells were determined by DCFDA assay. Paired t-test p values; *p < 0.05, **p < 0.01, ***p < 0.001.
