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. 2001 May 1;98(10):5786–5791. doi: 10.1073/pnas.101116298

Figure 5.

Figure 5

Different AP-1 compositional patterns after UVB stimulation and Inh I or Inh II treatment as assessed by gel supershift assays. Cl 41 cells were cultured and treated, nuclear proteins were isolated, and electrophoretic mobility-supershift assays were carried out as described in the text. Negative control (lane 1) indicates untreated cells. Rabbit serum (RS) was used as an internal control because the antibodies used for the supershift assay were dissolved in rabbit serum. Arrows indicate supershifted bands. (A) Cl 41 cells without any treatment. Incubation of extracted nuclear proteins with JunD or Fra-2 antibodies induced a clearing of the AP-1 band or a supershifted band (lanes 5 and 9). (B) Cl 41 cells were treated with UVB. Incubation of extracted nuclear proteins with c-Jun or c-Fos antibody induced a strong supershifted band (lanes 4 and 7). Incubation of extracted nuclear proteins with JunB antibody also induced a slight supershifted band (lane 5). In contrast to unstimulated control cells (A), incubation of nuclear extracts with JunD or Fra-2 antibodies failed to induce clearing of the AP-1 band or a supershifted band (lanes 6 and 10). (C) Cells were treated with Inh I followed by exposure to UVB. Incubation of extracted nuclear proteins with JunD or Fra-2 antibodies once again induced a clearing and supershifted band (lanes 5 and 9). The bands induced by c-Jun or JunB antibodies can still be seen but with attenuation (lanes 3 and 4). (D) Treatment with Inh II followed by exposure to UVB shows results similar to those seen with Inh I. Rabbit serum did not induce any supershifted bands in these experiments (lane 2 of A and lane 3 of B).