Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of RPE-1 nuclei mock treated and treated with myriocin for 24 h. Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. (B) Percentage of nuclear abnormalities of RPE-1 cells treated with myriocin (n = 100 cells). (C) Representative images of RPE-1 nuclei upon knockdown of SPTLC1 or SPTLC2 after 72 h. (D) Percentage of nuclear abnormalities of RPE-1 cells upon knockdown of SPTLC1 or SPTLC2 (n = 100 cells). (E) Western blot analysis of SPTLC1 and SPTLC2 protein levels in RPE-1 cells. (F) Representative images of RPE-1 nuclei treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (Cer) for 24 h. (G) Percentage of RPE-1 cells showing abnormal nuclear morphology when treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (n = 100 cells). (H) Representative images of primary human skin fibroblast (HSF) nuclei mock treated and treated with myriocin for 24 h. (I) Percentage of nuclear abnormalities of primary HSF treated with myriocin (n = 100 cells). (J) Representative images of primary HSF nuclei treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide for 24 h. (K) Percentage of primary HSF showing abnormal nuclear morphology when treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (Cer) (n = 100 cells). All images are representative of three biological replicates. Scale bars (A, C, F, H, and J), 10 μm.

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Staining, Knockdown, Western Blot

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of primary human fibroblasts from 2 euploid donors and 2 patients with Down syndrome. Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. Scale bar, 2.5 μm. (B) Percentage of fibroblasts from euploid donors showing abnormal nuclear morphology when treated with fumonisin B 1 or SKi-II (n = 100). (C) Percentage of fibroblasts from 2 patients with Down syndrome showing abnormal nuclear morphology treated with fumonisin B 1 , SKi-II, or ceramide (Cer) (n = 100). (D) Representative images of trisomy 21 nuclei treated with fumonisin B 1 , SKi-II, or ceramide. Scale bar, 10 μm. (E) Representative images of primary human fibroblasts from 2 euploid donors and 2 patients with Patau syndrome(trisomy 13) or Edward syndrome (trisomy 18). Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. Scale bar, 2.5 μm. (F) Percentage of fibroblasts from 2 patients with Patau syndrome showing abnormal nuclear morphology treated with fumonisin B1, SKi-II, orceramide (n = 100). (G) Percentage of fibroblasts from 2 patients with Edward syndrome showing abnormal nuclear morphology treated with fumonisin B1, Ski-II, or ceramide (n = 100).

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Staining

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of primary human fibroblasts from euploid donors and patients with Patau syndrome (trisomy 13), Edward syndrome (trisomy 18), or Down syndrome (trisomy 21). Immunofluorescence is shown for lamin A/C, H3K9triMe, and Hoechst 33342. In the merge images, lamin A/C is green, H3K9triMe is red, and DNA is blue. Scale bar, 5 μm. (B) Western blot analysis of H3K9triMe in human fibroblasts. (C) Quantification of the number of 53BP1 foci in human fibroblasts mock treated or in the presence of fumonisin B 1 (n = 100). (D) Representative images of primary human fibroblasts from euploid donors and patients with Down syndrome. Immunofluorescence is shown for 53BP1 (red) and Hoechst 33342 (blue). There is no correlation between abnormal nuclear morphology and number of 53BP1 foci. Scale bar, 5 μm. (E) Growth curves of primary human fibroblasts from 2 euploid donors and 2 patients with Patau (trisomy13), Edward (trisomy18), or Down syndrome(trisomy 21) with increasing concentrations of fumonisin B 1 . (F) Representative images of primary human astrocytes from 4 euploid donors and 4 donors with Down syndrome. Immunofluorescence for the nucleus is blue stained with Hoechst 33342. Scale bar, 5 μm. (G) Percentage of astrocytes from euploid and trisomy 21 donors showing abnormal nuclear morphology (n > 200).

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Western Blot, Staining

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet:

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Recombinant, Membrane, Protease Inhibitor, Control, Software