Fig. 4.
AMBRA1-RFP-sspB-mediated mitophagy can be induced in T lymphocytes. a Human T lymphocytes from healthy donors were double infected with viral vectors encoding Venus-iLID-ActA/AMBRA1-RFP-sspB or Venus-iLID-ActA/RFP-sspB (negative control). Cells were subsequently illuminated 24 h with pulsed (1 s light, 1 min dark) blue light or kept in the dark, then fixed and immunostained for Tom20 (cyan). Nuclei were counterstained with DAPI (blue). The graph show the intensity per cell of the Tom20 signal from four different donors. A minimum of 50 cells were analyzed per donor. Scale bar: 10 μm. Data shown: mean ± S.E.M. Hypothesis test: ANOVA test. #p < 10−4. n.s. not statistically significant. b Human T lymphocytes, manipulated as described in (a), were lysed and analyzed by WB. Tom20 and SOD2 were used as mitochondrial markers, while GAPDH was the loading control. AMBRA1-RFP-sspB was detected to verify the rate of overexpression. Graphs recapitulate the normalized ratio between the densitometric signals of the two mitochondrial markers over GAPDH in four different donors. Data shown: Mean ± S.E.M. Hypothesis test: ANOVA test. *p < 5 × 10−2. Mr (kDa): Relative molecular mass expressed in kilodalton. Source data are provided as a Source Data file