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. Author manuscript; available in PMC: 2015 Sep 15.
Published in final edited form as: Arch Biochem Biophys. 2014 Jul 5;558:143–152. doi: 10.1016/j.abb.2014.06.021

Figure 3. EISO inhibited UV-induced AP-1 activity without affecting signaling pathways upstream of AP-1 known to be induced by UV.

Figure 3

(A) HCL-14 cells, a HaCaT cell line stably expressing a portion of the human collagenase I gene (containing an AP-1 binding site) driving a firefly luciferase reporter gene, were treated with EISO for 1 hr prior to UV irradiation. After exposure to UV, media containing the same pretreatment (DMSO or EISO at the indicated concentration) was added back to the cells for 12 hr before cell lysates collected. Luciferase activity was measured and normalized to total protein content. Data are expressed as percent of the UV-induced luciferase activity in DMSO (vehicle)-treated control cells. (B) HaCaT cells were pretreated with EISO for 1 hr, irradiated with UVB and then returned to media containing pretreatments for 0.5 or 1 hr. Cell lysates were subjected to Western analysis using antibodies against phospho-Akt, phospho-JNK, phospho-ERK 1/2, and phospho-p38. Total ERK 1/2 was also analyzed to show that equal protein was loaded on the gel across all treatments. (C) FL-30 cells, a HaCaT cell line that stably expresses a full-length human c-fos promoter driving a firefly luciferase reporter gene, were pretreated with EISO for 1 hr before irradiation with UV and then returned to media containing pretreatments. Cell lysates were collected 6 hr post UV-irradiation and analyzed for luciferase activity, which was normalized to total protein.