Figure 2 from Claudin-4 Modulates Autophagy via SLC1A5/LAT1 as a Mechanism to Regulate Micronuclei

Claudin-4 links genomic instability to autophagy in HGSOC cells. We analyzed the functional role of claudin-4, establishing a link between autophagy and micronuclei (indicator of genomic instability) in HGSOC cells. This analysis was conducted both in vitro (HGSOC-GFP-mCherry-LC3 expressing cells) and in vivo (PDX in a humanized mice model). The assessment involved techniques such as flow cytometry, confocal microscopy (CM), and multispectral immunofluorescence. A–C, show percentage autophagy flux in HGSOC cells (flow cytometry) during CMP treatment (400 µmol/L), and claudin-4 genetic manipulation at 24 and 48 hours of culture (four independent experiments; Dotted blue triangles suggest an increase in autophagy). D–F, show confocal images of HGSOC cells expressing GFP-mCherry-LC3 and stained with DAPI (white arrow indicates the convergence of autophagy flux (mCherry) with micronuclei. G, Top, shows confocal images (from live-cell imaging) highlighting (yellow dotted square) a micronucleus; (bottom), kymographs generated from live-cell confocal imaging (from yellow dotted square). It shows the mobility pattern over time of the same region in every channel (Hoechst, GFP, and mCherry). H, Shows a line scan that indicates the fluorescence pattern for the micronucleus and autophagosomes (mCherry) from (G). (significance, P < 0.05). Graphs show mean and SEM, scale bar, 10 and 5 µm.