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. 2018 Mar 26;7:e34122. doi: 10.7554/eLife.34122

Figure 2. D1/HMGA1 loss-of-function results in micronuclei formation, and defective nuclear envelope integrity.

(A, B) Control (D1LL03310/+) (A) and D1LL03310/Df mutant (B) spermatogonial cells stained for DAPI (red), Vasa (blue) and LaminDm0 (green). Arrow indicates micronucleus. Bars: 5 μm. (C) Quantification of micronuclei-containing cells from +/+ control (n = 269) and D1LL03310/Df (n = 334) from three independent experiments. p-Value from student’s t-test is shown. Error bars: SD. (D, E) siControl (D) and siHMGA1 transfected (E) MOVAS cells stained for DAPI (blue), HMGA1 (red) and Lamin (green). Arrow indicates micronucleus. (F) Quantification of micronuclei-containing cells in siControl (n = 518) and siHMGA1 (n = 588) transfected cells from four independent experiments. (G, H) Control (D1LL03310/+) (G) and D1LL03310/Df (H) spermatogonia expressing nls-GFP (green) stained for Vasa (blue) and LaminDm0 (red). nlsGFP was observed in cytoplasm in D1LL03310/Df spermatogonia. (I) Quantification of spermatogonia with cytoplasmic GFP (>1 μm exclusions or pan-cytoplasmic) in D1LL03310/+ (n = 810) and D1LL03310/Df (n = 780) testes from two independent experiments. (J, K) D1LL03310/+ (J) and D1LL03310/Df (K) spermatogonia expressing ER-GFP marker (green) stained for Vasa (blue) and LaminDm0 (red). Arrowhead points to ER marker-positive micronucleus. Arrows point to site of weak nuclear LaminDm0 staining. (L, M) Control (D1LL03310/+) (L) and D1LL03310/Df (M) spermatogonia stained for Vasa (blue) and LaminDm0 (green) and Otefin (red). Arrowhead points to Otefin-containing micronucleus. Arrows point to site of weak nuclear LaminDm0 staining.

Figure 2.

Figure 2—figure supplement 1. Formation of micronuclei upon chromocenter disruption in C3H10T1/2 and C2C12 mouse cells.

Figure 2—figure supplement 1.

(A, B) siControl (A) and siHMGA1 (B) transfected C3H10T1/2 cells stained for DAPI (blue), HMGA1 (red) and LaminB (green). Arrowhead indicates micronuclei. Bars: 5 μm. (C) Quantification of micronuclei containing cells from siControl (n = 291) and siHMGA1 (n = 303) transfected cells from three independent experiments. p-Value from student’s t-test is shown. Error bars are SD. (D, E) siControl (D) and siHMGA1 (E) transfected C2C12 cells stained for DAPI (blue), HMGA1 (red) and LaminB (green). Arrowhead indicates micronuclei. Bars: 5 μm. (F) Quantification of micronuclei containing cells from siControl (n = 953) and siHMGA1 (n = 699) transfected cells from three independent experiments. p-Value from student’s t-test is shown. Error bars are SD.