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. 2016 Jan 9;7(8):8916–8930. doi: 10.18632/oncotarget.6866

Figure 2. Gene 33 expression is regulated by Cr(VI) through multiple mechanisms.

Figure 2

A. BEAS-2B cells were either left untreated or treated with 5 μM Cr(VI), with or without presence of 20mM NAC for 24 or 48 hours. Cells were then harvested and Western blotted with antibodies toward the indicated proteins. B. BEAS-2B cells where left untreated or treated with 10μM MG132, 5 μM Cr(VI), or 10μM MG132 plus 5 μM Cr(VI) for 24 or 48 hours as indicated (MG132 was added 6 hours before the end of the treatment). Cells were then processed as in A. C. BEAS-2B cells were co-transfected with plasmids containing FLAG-tagged Gene 33 and FLAG-tagged ubiquitin. Twenty fours later, cells were left untreated or treated with 5 μM Cr(VI), with or without presence of 10 μM MG132. Cells were then processed as in A. D. Total cellular proteins from C were Western blotted with an antibody toward FLAG. J. BEAS-2B cells were transfected with an miRNA mimics (Ctrl) or an miR200c inhibitor (200c). Seventy hours after transfection, the cells were treated with or without Cr(VI) for 48hours. The Gene33 expression levels were then determined by Western blot. Data are presented as mean +/− SD (E,F,H,I).