Fig. 2: Impaired chromosome reorganization upon VRK-1 depletion is mediated by BAF-1. | Nature Communications

Fig. 2: Impaired chromosome reorganization upon VRK-1 depletion is mediated by BAF-1.

From: BAF-1–VRK-1 mediated release of meiotic chromosomes from the nuclear periphery is important for genome integrity

Fig. 2

a Insets from vrk-1::AID::ha germlines stained with DAPI (white) and anti-PiSer8 SUN-1 antibody (magenta) after depletion of VRK-1, baf-1RNAi, or both. Magenta arrows delineate meiotic entry. Scale bar: 10 µm. b Percentage of clustered chromatin nuclei in the first 10 meiotic cell rows in the vrk-1::AID::ha germlines. Mean ± SD, L4440 (empty vector)–ethanol: 41.0% ± 6.5%; n = 6 vs L4440–auxin: 21.5% ± 7.9%; n = 8, P < 0.0001 Two-sided Fisher’s exact test; baf-1 RNAi–ethanol: 45.7% ± 12.1%; n = 5, P = 0.3663, Fisher’s exact test compared with L4440–ethanol; baf-1 RNAi–auxin: 45.9% ± 6.9%; n = 8, P = 0.3103, Fisher’s exact test compared with L4440–ethanol. n= number of germlines. c DAPI staining (white) and PiSer8 SUN-1 immunostaining (magenta) in -1 oocytes of vrk-1::AID::ha germlines. Scale bar: 10 µm. d Transition zone insets from vrk-1::AID::ha; flag::baf-1 germlines treated with ethanol (top) or auxin (bottom) and immunostained for FLAG (yellow) and co-stained with DAPI (white). Scale bar: 10 µm. e Average line profile analysis of FLAG signal intensity in the center of the nucleus for the indicated treatments: ethanol, n = 63 nuclei for progenitor zone, 71 for transition zone; and auxin, n = 94 for progenitor zone, 93 for transition zone. f Transition zone insets from flag::baf-1, flag::baf-1T3A, flag::baf-1S4A germlines immunostained for PiSer8 SUN−1 (magenta) and co-stained with DAPI (white). Scale bar: 10 µm. g Percentage of clustered chromatin nuclei in the first 10 meiotic cell rows of germlines in the indicated genotypes. Mean ± SD, flag::baf-1: 52.3% ± 10.8%, n = 9 germlines; flag::baf-1T3A: 47.4% ± 12.8%, n = 8 germlines; P = 0.1217; flag::baf-1S4A: 17.9% ± 11.5%, n = 8 germlines; P < 0.0001 two-sided Fisher’s exact test). h Western blot of the indicated cellular fractions from vrk-1::AID::ha worm pellets for the indicated genotypes treated with ethanol or auxin. Arrows indicate the presence (in ethanol) or absence (in auxin) of the BAF-1 S4Pi band. Full blots included in Source table. Anti-GAPDH and anti-H3 antibodies were used as loading controls. GAPDH = glyceraldehyde-3-phosphate dehydrogenase, H3 = Histone 3, kDA = KiloDalton.

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