32 hpfs  (ScienCell)

Journal: PLoS ONE

Article Title: Embryonic Senescence and Laminopathies in a Progeroid Zebrafish Model

doi: 10.1371/journal.pone.0017688

Figure Lengend Snippet: ( A ) Muscle fiber visualization in zlamin A/C-knockdown embryos. (a–d) The left panels show a lateral view of the entire embryo bodies and a magnified bright field view of the somites. (e–h) Lateral views of MO-injected embryos at 24 hpf stained with the F59 antibody (adaxial cells/slow muscle fibers). In panel (f), white arrowheads indicate broken or missing muscle fibers. In panel (h), white arrowheads indicate the wavy muscle fibers in compared with control embryos. (i–l) The right panels show cryostat sections of 48 hpf LMNA -MO injected embryos stained for fast muscle fibers (F310). ( B ) Percentage of muscle phenotypes scored for MO-injected embryos at 24 hpf. The graph shows the distribution of phenotypes observed following injections with 4 ng of MOs. Migration failure in embryos is represented in gray and wavy muscle fibers in black. The percentage of each muscle phenotypes ‘waving’, ‘migration failure’, ‘normal’ were, respectively, MO1-5mis; 7.5%, 5.0%, 87.5%; MO1-MO; 3.8%, 46.2%, 50.0%; MO2-5mis; 6.1%, 6.1%, 87.8%; MO2-MO; 29.4%, 25.5%, 45.1%. ( C ) Cartilage in 6 dpf larvae were stained with Alcian blue and then whole-mounted. (a–d) Lateral views of the head portions of living larvae are shown in the top panels. (e–h) Lateral view of Alcian blue stained larvae. (i–l) Ventral views of Alcian blue stained larvae. The red lines indicate the ceratohyal articulates (ch), and the red arrow indicates the Meckel's (m) to palatoquadrate (pq).

Article Snippet: The following primary antibodies were used for immunostaining: 24 hpf embryos for F59 (1∶100) and 48 hpf embryos for F310 (1∶100) (DSHB, Iowa City, IW).

Techniques: Knockdown, Injection, Staining, Control, Migration

Journal: PLoS ONE

Article Title: Embryonic Senescence and Laminopathies in a Progeroid Zebrafish Model

doi: 10.1371/journal.pone.0017688

Figure Lengend Snippet: ( A ) External features of z progerin transgenic zebrafish (30 hpf) without (Control, -FTI) or with FTI treatment (100 µM, +FTI). ( B ) Amelioration of nuclear morphology by FTI treatment (100 µM, +FTI) in comparison with untreated (-FTI) z progerin fish. Analysis of the misshapen nuclear morphology was also performed by using different concentrations of FTI in z progerin transgenic embryos at 30 hpf as shown in right graph. The number of cells with abnormally shaped nuclei were determined by fluorescence microscopy (* P <0.05). ( C ) Amelioration of muscle phenotype (stained with the F59 antibody) at 24 hpf and reduction of SA-β-gal activity at 3.5 dpf in z progerin fish embryos by FTI treatment (100 µM, +FTI) in comparison with the untreated condition (-FTI). ( D ) Western blot analysis of prelamin A accumulations in AB9 zebrafish fibroblasts following FTI treatment, and the quantitation of prelamin A amounts in right graph. The quantifications of intrinsic prelamin A accumulations were shown are the averages from two-independent experiments.

Article Snippet: The following primary antibodies were used for immunostaining: 24 hpf embryos for F59 (1∶100) and 48 hpf embryos for F310 (1∶100) (DSHB, Iowa City, IW).

Techniques: Transgenic Assay, Control, Comparison, Fluorescence, Microscopy, Staining, Activity Assay, Western Blot, Quantitation Assay

Journal: PLoS ONE

Article Title: Embryonic Senescence and Laminopathies in a Progeroid Zebrafish Model

doi: 10.1371/journal.pone.0017688

Figure Lengend Snippet: Summary of distinct zebrafish models for laminopathy in this study.

Article Snippet: The following primary antibodies were used for immunostaining: 24 hpf embryos for F59 (1∶100) and 48 hpf embryos for F310 (1∶100) (DSHB, Iowa City, IW).

Techniques: Transgenic Assay, Expressing, Blocking Assay, TUNEL Assay