Journal: bioRxiv

Article Title: Selective autophagy of whole micronuclei suppresses chromosomal instability

doi: 10.64898/2026.04.03.716211

Figure Lengend Snippet: a, Schematic of the H2B-mKeima chromatin acidification reporter. Primary excitation shifts from 488nm to 561nm laser upon acidification b, H2B-mKeima acidic to neutral signal ratio change in micronuclei upon acidification (n=30MN). c, Acidification rate of spontaneously arising MN in different cell lines (n between 20 and 50 MN per cell line per experiment, 3 separate experiments). d, Acidification rate of MN induced with different methods (n≥100MN per experiment, 3 separate experiments). e, Montage showing MN targeted by LC3B, engulfed in an autophagosome-like structure, followed by H2B-mKeima acidification (scale bar = 5μm). f, Montage showing MN capture within LC3B positive autophagosome, followed by lysosome fusion and reporter acidification (scale bar = 3μm). g, Quantification of LC3 recruitment on MN before reporter acidification (n=20 MN per fate). h, Holotomography montage showing membrane recruitment around MN before acidification of the reporter (scale bar = 2μm). i, MN acidification rates over the course of 14h after nocodazole shake-off upon silencing of main autophagic components (n≥100MN per experiment, 4 separate experiments). j, MN acidification rate in ATG7ko cells (n≥100MN per experiment, 4 different clones). k, MN acidification rates in cells treated with autophagy modulators (n≥100MN per experiment, 4 separate experiments). l, MN abundance in cells treated for 16h with autophagy modulators after nocodazole shake-off (n≥500 cells per experiment, 4 separate experiments). Data are shown as mean ± sem, and were analysed using a one-sample t test (c, l), two-way ANOVA (g, i, k), unpaired t-test (j) (* = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001)

Article Snippet: To investigate the impact of suppression of Aurora B midzone localisation on MN fates and Lamin B1 levels Paprotrain (10μM, MedChemExpress, HY-101298) was added from nocodazole washout or after mitotic exit.

Techniques: Membrane, Clone Assay

Journal: bioRxiv

Article Title: Selective autophagy of whole micronuclei suppresses chromosomal instability

doi: 10.64898/2026.04.03.716211

Figure Lengend Snippet: a, Montage showing BAF dissociation from the MN membrane before acidification and timecourse of BAF (cortical/inner) signal for MN populations with different fates (scale bar = 1μm). b, Fate of MN with sustained (≥ 3 consecutive frames) BAF dissociation (n≥50MN per experiment, 3 separate experiments). c, Fate of MN with sustained (≥ 3 consecutive frames) Lamin B1 dissociation (n≥50MN per experiment, 3 separate experiments). d, LC3 recruitment on MN after sustained BAF dissociation (n=22 MN from 2 individual experiments). e, Relation between initial import capacity and final BAF localization based on MN fate. f, Acidification rate of MN in cells treated with the MKLP2 inhibitor, paprotrain, from nocodazole washout (pre-anaphase) or in the next interphase (n≥100MN per experiment, 4 separate experiments). g, Acidification rate of MN from cells with or without Lamin B1 overexpression(n≥100MN per experiment, 4 separate experiments). h, Lamin B1 levels on lagging derived MN in cells treated with or without paprotrain from nocodazole washout (n=50 individual MN, 2 separate experiments). i, Acidification rate of MN in cells treated with the VRK1 inhibitor, VRK1-IN-1, from nocodazole washout (pre-anaphase) or in the next interphase (n≥100MN per experiment, 4 separate experiments). j, Timecourse of BAF localization in acidified MN in basal conditions or following VRK1-IN-1 treatment from nocodazole washout (n between 14 and 21 MN from n=2 experiments). k, Holotomography montage of adjacent MN (white arrow=acidification-fated; green arrow=stable) several hours (top) and shortly (bottom) before acidification, showing rapid changes in the morphology of the acidification-fated MN shortly before acidification (scale bar = 1μm). l, CLEM experiment showing deformed but intact NE of autophagy targeted MN (green = LC3 positive autophagosomal membrane; orange = intact NE) (scale bar = 2μm for immunofluorescence image and scale bar = 500nm for EM image). Data are shown as mean ± sem, and were analysed using a one-way ANOVA (b, g), unpaired t-test (c, h), one-sample t-test (d), two-way ANOVA (f, i) (ns = non-significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001)

Article Snippet: To investigate the impact of suppression of Aurora B midzone localisation on MN fates and Lamin B1 levels Paprotrain (10μM, MedChemExpress, HY-101298) was added from nocodazole washout or after mitotic exit.

Techniques: Membrane, Over Expression, Derivative Assay, Immunofluorescence