Figure 5.

DFCP1 is critically involved in HCV and SARS-CoV-2 replication

(A) DFCP1 KD in Huh7-Lunet/T7 cells was achieved by siRNA transfection (two different siRNAs) and confirmed by western blotting. GAPDH served as loading control.

(B and C) Huh7-Lunet/T7 cells were transfected twice with siRNAs using a 24-h interval, prior to electroporation with a subgenomic HCV reporter replicon 48 h after the second siRNA transfection.

(B) Cell viability (ATP content) was measured 24, 48, and 72 h after the last siRNA transfection.

(C) Luciferase activity reflecting HCV RNA replication was measured 24, 48, and 72 h after electroporation.

(D) DFCP1 KD in A549/ACE2 cells was evaluated by using western blotting as described for (A).

(E and F) A549/ACE2 cells were transfected with siRNAs 48 h prior to infection with SARS-CoV-2 (MOI of 1).

(E) Cell viability (ATP content) was measured 24 h after siRNA transfection.

(F) Relative SARS-CoV-2 replication was determined by N protein-specific immunostaining at 24 h post-infection and normalization of the values to those obtained with NT siRNA treated cells.

All data represent the mean ± SEM from three independent experiments.