Figure 4. Western immunological analysis of uninfected cells.

HeLa cells were grown to near confluence in the same medium that was used to perform the infection with Ct but cells were not infected. Panel A. Nuclear and cytosolic samples were obtained as described in the Method section. As confirmed by coomassie blue staining (right panel), an equal amount of proteins (10 µg) of the nuclear (lane N) and cytosolic (lane C) samples were loaded in each lane of a SDS-polyacrylamide 4–16 % gradient gels. After migration, the proteins were transferred on PVDF membranes that were blotted with antibody against ACBD6, ZNF23 or ANKRD7. ANKRD7 is a known nuclear protein and was used as a control [66]. The molecular mass standard (Precision Plus Protein Dual-Stained, Bio-Rad) is indicated on the right of the panels. A band of the predicted molecular mass of 32 kDa and 64 kDa was detected for ABCD6 and ZNF23 respectively, in the cytosolic and nuclear samples. A major band of the expected size of 30 kDa (asterisk) was detected for ANKRD7 in the nuclear extract. Panel B. ACSL3 protein was detected with the mouse anti-ACSL3 antibody in a total protein extract of HeLa cells as described in panel A.