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. 2023 Mar 13;13:1057082. doi: 10.3389/fcimb.2023.1057082

Figure 2.

Figure 2

Effect of HCV fitness on the level of cellular protein H3Ser10ph and viral proteins NS5A and core. (A) Huh-7.5 reporter cells were either mock-infected or infected with HCV p0, HCV p100 or HCV p200 (abbreviated as p0, p100 and p200, respectively) at an initial MOI of 0.03 TCID50/cell; protein extracts were prepared at the indicated times post-infection. Bands are those visualized by Western blot assays of cellular protein H3ser10ph and viral proteins NS5A and core (with β-actin as loading control). (B) H3Ser10ph levels in mock-infected or HCV-infected cells expressed as the fold induction relative to the corresponding value for the mock-infected cells; the infecting HCV (code in upper box), and the time post-infection at which extracts were prepared are given in the abscissa; the numerical densitometry values (measured relative to the mock-infected sample, taken as 1) are indicated in the upper box, next to the panel. Asterisks indicate statistical significance as follows: *p<0.05; **p<0,01; ***p<0.00; unpaired t-test. The values are the result of three independent experiments (biological triplicate). (C) Correlation between the increase of intracellular viral titer (code of infecting virus in upper box) expressed as log. 10 TCID50/ml/h [data are from (28) and they are indicated in the box above the panel, for mock, HCV p0, HCV p100 and HCV p200, respectively] and the relative H3Ser10ph densitometry values at time 72 hours post-infection (given in panel B). The discontinuous line corresponds to function y=-3.45x+0.81 (R2 = 0.77, p-value=0.1227; linear regression test). (D) Same as B but for viral protein NS5A. (E) Same as C but for the correlation between viral protein NS5A and cellular protein H3Ser10ph levels (densitometry values in panels B and D). The discontinuous line corresponds to function y=-1.33x+1,13 (R2 = 0.77; p-value=0.00018; linear regression test). (F) Same as B but for viral core protein. (G) Same as C but for the correlation between viral core protein and H3Ser10 levels (densitometry values in panels B and F). The discontinuous line corresponds to function y=-2.13x+1.12 (R2 = 0.85; p-value=0.000018; linear regression test).