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. 2012 Aug 7;3(8):798–810. doi: 10.18632/oncotarget.574

Figure 5. Fibroblasts over-expressing MFF show increased ROS generation, and induction of autophagy, with NF-kB hyper-activation.

Figure 5

(A) ROS levels were measured by FACS analysis, as described under Materials and Methods. Note that MFF over-expression induces a 50% increase in ROS production, as compared to control cells. (B) Induction of an autophagic/mitophagic process was analyzed on cellular extracts of fibroblasts expressing MFF and empty vector controls by immunoblotting. Note that MFF-fibroblasts show the up-regulation of several markers of autophagy (LC3I/II and cathepsin B), and mitophagy (BNIP3 and BNIP3L), as compared to control cells. No changes were observed for Beclin-1 expression. β-actin is shown as an equal loading control. (C) NF-kB activation was monitored by immuno-blotting with phospho-NF-kB antibodies. Note that MFF-fibroblasts show constitutive NF-kB activation, relative to control cells, suggesting that NF-kB activation maybe the mechanism driving the induction of autophagy. β-actin was used as an equal protein loading control.