Table 1.
List of Primer sequences for cloning.
Table 2.
List of primers for the selected genes.
Fig 1.
Confirmation of P2X4 gene in vector pcDNA3.1+ Marker: λHindIII digested marker.
Lanes 4, 10, 11, and 13 showing digested plasmid with the release of insert.
Fig 2.
Confirmation of P2X4 gene in vector pcDNA3.1+ pcDNA 3.1+ mammalian expression vector (5.0-kb), Insert P2X4 (1.7-kb).
Fig 3.
Map of construct pcDNA3.1+/P2X4.
Fig 4.
Plasmid pcDNA3.1+/P2X4 restriction digestion after mini prep.
Lane 1 showed: λHindIII digested marker, Lanes 2 and 3 showing digested plasmid with release of insert, pcDNA 3.1+ mammalian expression vector (5.0-kb), Insert P2X4 (1.7-kb).
Fig 5.
Linearization of vector pcDNA3.1+/P2X4 before transfection.
Lane 1 showed: λHindIII digested marker, Lanes 4, 5, 6 and 7 showing digested linerized plasmid pcDNA3.1+/P2X4 (6.7-kb).
Fig 6.
RT-PCR of cell line 293T/P2X4.
Lane 1 showed: 50-bp DNA size marker, Lanes 2 &3 showing amplified product of transfected P2X4 (390-bp).
Fig 7.
Lane 1 showed: 50-bp DNA size marker, Lanes 2,3.4 and 5 showing amplified product for NV (180-bp) (T7and BGH).
Fig 8.
RT-PCR of single clones of cell lines 293T/P2X4 and 293T/NV.
Lane 1 showed: 100-bp DNA size marker, Lanes 2 &3 showing amplified product of NV (180-bp), Lanes4-15 showed amplified product of transfected P2X4 (390-bp).
Fig 9.
Protein expression analysis of P2X4 receptor.
Western blot of P2X4 receptor (Protein lysate of selected single clones of cell lines 293T/P2X4 and 293T/NV). The predicted molecular mass of P2X4 is ~ 46kDa, but it is generally detected near 60–70 kDa because of glycosylation.
Fig 10.
HCV RNA was quantified by RT-QPCR after infection in stable cell lines.
All values are expressed as mean ± SEM *P≤0.05 vs. control NV/HCV.
Fig 11.
Expression of P2X4 gene at day 5 of post infection with HCV.
All values are expressed as mean ± SEM, *P≤0.05 vs. control P2X4-NR cells.
Fig 12.
Expression of P2X4 gene at day 9 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control P2X4-NR cells.
Fig 13.
Expression of antioxidants on day 5 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control NV/HCV.
Fig 14.
Expression of antioxidants on day 9 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control NV/HCV.
Fig 15.
Expression of various cytokines on day 5 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control NV/HCV.
Fig 16.
Expression of various cytokines on day 9 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control NV/HCV.
Fig 17.
Expression of ECM markers on day 5 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control.
Fig 18.
Expression of various cytokines on day 9 of post infection.
All values are expressed as mean ± SEM, *P≤0.05 vs. control NV/HCV.
Fig 19.
Schematic overview of the purinergic signaling in the presence of HCV.
HCV interrupts purinergic signaling Increased ATP in the extracellular mileu increases oxidative stress and fibrogenic cytokines and ECM cytokines leading towards fibrosis.