Fig. 4
AFB1 inhibits STAT1 protein synthesis. HepG2 cells were cultured at density of 5 × 104 cells per well of the 6-well plate until they reached 80% confluence. The cells were stimulated and treated as described above. After blotting, the membranes were immunoblotted with antibody against the STAT1 epitope of the STAT1 protein as primary antibody followed by horseradish peroxidase conjugated goat anti-rabbit antibody as secondary antibody. The membranes were also probed for GAPDH which was used as endogenous control. The dilutions of the primary antibodies used against STAT1 and GAPDH proteins were as follows: STAT1 (Thermo Scientific, cat no PA5-34504; 1:1000), GAPDH (Thermo Scientific, cat no QE212271; 1:2500). The secondary antibody used was polyclonal goat anti-Rabbit conjugated to HRP (Thermo Scientific, cat no PI31460, 1: 5000). The blotted membranes were developed using enhanced chemiluminescence reagents and the protein bands were detected and captured with C-DIGIT blot scanner imaging device. (a) The relative STAT1 protein intensity was quantified using Image J software package. (b) Western blot analysis of STAT1 protein. (c) Western blot analysis of GAPDH protein as loading control