Research Paper Volume 13, Issue 1 pp 957—972

Bnip3 interacts with vimentin, an intermediate filament protein, and regulates autophagy of hepatic stellate cells

Bnip3 was partially co-localized with autophagosomes in activated HSCs and inhibition of Bnip3 led to the blockage of the autophagic flow. LX-2 cells were treated with 100 μM CoCl2 or 2 μg/ml LPS for 8 h. Culture-activated primary HSCs from mice were cultured up to 2 days or 6 days. Immunofluorescence assay was performed to detect Bnip3 (Cy3) and LC3B (FITC) in LX-2 cells (A) and culture-activated primary HSCs from mice (B) by confocal microscopy. (C) Primary HSCs were isolated from mice and seeded in coverslips. Cells were transfected with specific siRNA targeting Bnip3 as cells were cultivated up to day 2 and GFP-RFP-LC3B plasmid was transfected into cells 24h later. Cells were fixed at day 6 and images were captured by confocal microscope to observe the formation of autophagosomes.

Figure 5. Bnip3 was partially co-localized with autophagosomes in activated HSCs and inhibition of Bnip3 led to the blockage of the autophagic flow. LX-2 cells were treated with 100 μM CoCl2 or 2 μg/ml LPS for 8 h. Culture-activated primary HSCs from mice were cultured up to 2 days or 6 days. Immunofluorescence assay was performed to detect Bnip3 (Cy3) and LC3B (FITC) in LX-2 cells (A) and culture-activated primary HSCs from mice (B) by confocal microscopy. (C) Primary HSCs were isolated from mice and seeded in coverslips. Cells were transfected with specific siRNA targeting Bnip3 as cells were cultivated up to day 2 and GFP-RFP-LC3B plasmid was transfected into cells 24h later. Cells were fixed at day 6 and images were captured by confocal microscope to observe the formation of autophagosomes.