FOSB proteins in the human orbitofrontal and dorsolateral prefrontal cortices (OFC and DLPFC, respectively). (a) Schematic picture of FosB and ΔFosB. The epitopes recognized by the three different antibodies tested are depicted. (b) Immunoblots made with HeLa cell and human OFC tissue extracts, and probed with ab11959 and LS-C528. Lanes 1 and 5:5 μg of protein from total extracts of Mock-transfected HeLa cells; lanes 2 and 6:5 μg of protein from total extracts of HeLa cells transfected withYOF235 (plasmid which contains a FosB insert); lanes 3 and 7: 1 μg of protein from total extracts of HeLa cells transfected with YOF254 (plasmid which contains a ΔFosB insert); lanes 4 and 8; 80 μg of protein from total extracts of OFC tissue. (c) Immunoblots made with HeLa cell and human OFC tissue extracts, and probed with sc-48 preincubated with or without blocking peptide. Lane 1: see (b), lanes 1 and 5; lanes 2 and 3: see (b), lanes 2 and 6; lanes 4 and 5: see (b), lanes 3 and 7; lanes 6 and 7: see (b), lanes 4 and 8. (d) Upper panels: images of blots made with serial dilutions of a reference sample (10–120 μg of protein per well). Dashed areas: bands which optical densities (ODs) were linearly dependent on MemCode™ OD as is shown in the lower panels. Lower panels: linear ranges of protein ODs (R² > 0.8 in all cases). AU, arbitrary units. (e) Images of immunoblots made with 100 μg of protein from total OFC and DLPFC tissue extracts. Lane 1:see (b), lanes 3 and 7); lane 20: see (b), lanes 2 and 6. IC, internal control; 1C, control number 1; 1A, alcoholic number 1; etc. (f) Comparison of the immunoreactivities of the 39 and 46 kDa forms of FOSB and the 37 kDa form of ΔFOSB in the OFC and DLPFC between human controls and alcoholics. Horizontal bars represent group means and standard error of the mean