Autophagy Is an Innate Mechanism Associated with Leprosy Polarization
Fig 6
Differential regulation of BECN1 and BCL2 proteins in skin lesion cells of T-lep and L-lep patients.
(A) Protein contents from tuberculoid (T-lep) and lepromatous (L-lep) lesion cells were analyzed by western blot with anti-BECN1 and anti-BCL2 antibodies. GAPDH antibody was used to verify protein amount loading. Densitometric band-intensity analysis of the blots was realized and the BECN1/GAPDH (T-lep, n = 6; L-lep, n = 5) and BCL2/GAPDH (T-lep, n = 4; L-lep, n = 4) ratios were expressed as arbitrary units (AU). Bars represent the mean values ± SEM. *P < 0.05, Mann-Whitney test. (B) Macrophages (MΦs) were isolated from skin lesions of T-lep and L-lep patients and cultured for 7 days in full medium. Cells were fixed and labeled for BECN1 (red), BCL2 (green) and DAPI (blue). Cytoplasmatic BCL2 colocalizes with BECN1 in L-lep MΦs, but not in T-lep MΦs. Colocalization profiles between cytosolic BCL2 dots and BECN1 were quantified and expressed as percentage of cell area. The expression of nuclear compartment-associated BCL2 was excluded from the analysis. Results represent the mean ± SEM from one of three immunofluorescence experiments that yielded similar results. **P < 0.01, Mann-Whitney test. Scale bar: 10 μm.