Figure 1 from Claudin-4 Modulates Autophagy via SLC1A5/LAT1 as a Mechanism to Regulate Micronuclei

Characterization of micronuclei during claudin-4 manipulation in HGSOC cells. HGSOC cells were treated with CMP (400 µmol/L; 48 hours). Then, cells were fixed and stained with DAPI to mark DNA and the components of the nuclear lamina, lamin B1 and lamin A/C. Subsequently, a morphometric characterization was performed through confocal microscopy. A, Illustration indicating the histological subtype of EOC cells used as in vitro system, as well as certain genetic alterations. B, (Left), frequency of micronuclei during claudin-4 overexpression; (right), confocal images (maximum projections) highlighting (dotted squares) micronuclei. Similar data is presented for claudin-4 downregulation in OVCA429 (C) and OVCAR3 cells (D). (n = 4,134 OVCAR8 cells; n = 3,315 OVCA429 cells; n = 4,653 OVCAR3 cells; Two-tailed Mann–Whitney test). E, Selected confocal images (OVCAR8 WT cells treated with the vehicle from (B) showing lamin B1 and lamin A/C. It is highlighted the lack of nuclear lamina in some micronuclei. Similar data is presented in (F and G) for OVCA429 and OVCAR3 cells, respectively. (three independent experiments; Kruskal–Wallis test with Dunn's multiple comparisons, P < 0.05). Graphs show mean and SEM, scale bar, 10 µm.