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

Journal:

Article Title: Accumulation of mutant lamin A causes progressive changes in nuclear architecture in Hutchinson-Gilford progeria syndrome

doi: 10.1073/pnas.0402943101

Figure Lengend Snippet: Changes in nuclear architecture as HGPS cells age in culture. Typical nuclei from HGPS cells in passages 6 (a), 13 (b), and 26 (c), normal aged human AG09602B (d), and foreskin fibroblast (e) controls after labeling with LA Ab. Nuclei in control cells appeared similar from passage 6 through passage 17. The lamina increased in prominence or thickness in HGPS cell nuclei by passage 26 (c). Earlier passage HGPS (a) and control cell nuclei (d and e) were normal in appearance. Passage 6 and 26 HGPS cells were double labeled with LA (g and j) and LB (h and k) Abs. Note the extensive coincidence in the staining patterns in the merged image at passage 6 (i) and the decrease in this coincidence in a cell by passage 26 (l). The table shows that the contour ratio decreased as a function of passage number in HGADFN003 cells. P values were calculated relative to passage 2 of the AG09602B fibroblasts. There was no significant change in the contour ratio in control cells between passages 2 and 17. In HGADFN003 whole-cell extracts immunoblotted with LA/C Abs, a band migrating between LA (upper band, *) and lamin C (lower band, *) was observed and became more prominent as the passage number increased (f). (Scale bars = 5 μm.)

Article Snippet: Fibroblasts from unaffected older adults (AG09602B derived from a 92-yr-old female; Coriell) and unaffected human infant foreskin fibroblasts (Coriell) were used as controls.

Techniques: Labeling, Staining

Journal:

Article Title: Accumulation of mutant lamin A causes progressive changes in nuclear architecture in Hutchinson-Gilford progeria syndrome

doi: 10.1073/pnas.0402943101

Figure Lengend Snippet: Pre-LA accumulation in the nuclei of HGPS cells. In passage 6 and 13 HGPS cells, the pre-LA pattern consisted of weakly staining nucleoplasmic foci (a and b) similar to control AG09602B fibroblasts (d). However, by passage 26, the pre-LA staining was much more intense in the lobulated nuclei and was primarily associated with the lamina region (c). (e–g) Electron microscopic observations of passage 26 HGPS HGADFN003 cells (e and f) and normal human foreskin fibroblasts (g). A high-magnification view of the nuclear envelope in a normal human foreskin fibroblast showed a normal array of heterochromatin adjacent to the nuclear envelope, making any lamina structure difficult to detect (g). A low-magnification view of a passage 26 HGADFN003 nucleus showed extensive lobulation (e). A higher-magnification view of a passage 26 cell showed a loss of peripheral heterochromatin and a prominent electron-dense lamina region associated with the inner nuclear envelope membrane (f). In f and g, the nucleus is to the left. [Scale bars = 5 μm (a–e) and 200 nm (f and g).]

Article Snippet: Fibroblasts from unaffected older adults (AG09602B derived from a 92-yr-old female; Coriell) and unaffected human infant foreskin fibroblasts (Coriell) were used as controls.

Techniques: Staining

Journal: iScience

Article Title: Supernumerary proteins of the human mitochondrial ribosomal small subunit are integral for assembly and translation

doi: 10.1016/j.isci.2024.110185

Figure Lengend Snippet: Stability of mS37 is regulated by CX 9 C motif (A) Location of mS37 within the 55S mitoribosome. (B) Conservation of the CHCHD4/MIA40-oxidizable twin CX 9 C motifs in mS37 and (C) subsequent folding. (C) Schematic representation of 4 mutated cysteine residues into serine (C1234S). (D) Immunoblot of FLAG-tagged wild-type and C1234S mS37. Asterisks indicate low levels of C1234S mS37. (E) Immunoblot of mS37 protein level in patient-derived fibroblasts.

Article Snippet: Human diploid male control fibroblasts , Jackson lab , N/A.

Techniques: Western Blot, Derivative Assay