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. 2010 Feb 23;5(2):e9367. doi: 10.1371/journal.pone.0009367

Figure 6. Assessing the induction and inhibition of autophagy by WIPI-1 puncta formation analyses in DJ-1 WT and DJ-1 deficient MEF.

Figure 6

GFP-WIPI-1 puncta-formation analyses reveals that forced modulation of the autophagic activity is possible in DJ-1 deficient cells. WT (MEF DJ-1 +/+) (A) or DJ-1 KO MEF (MEF DJ-1 −/−) (B) were treated with standard inducers of autophagy, rapamycin and EBSS (medium lacking amino acids), a standard inhibitor of autophagy, wortmannin, or left untreated (mock treatment, control). Quantitative confocal microscopy was conducted. MEFs that displayed GFP-WIPI-1 puncta were counted as positive (puncta), and MEFs that did not displayed GFP-WIPI-1 were counted as negative (non-puncta). In total 200–300 MEFs were counted from 3 independent experiments (see supplemental figure S2A-H, supplemental table Table S1). P-values for puncta-positive DJ-1 WT cells upon rapamycin ( = 0,00067), wortmannin ( = 0,00026), EBSS ( = 0,00083), and p-values for puncta-positive for DJ-1 definient cells upon rapamycin ( = 0,00111), wortmannin ( = 0,00138), EBSS ( = 0,00117) are indicated (***).