Journal: Molecular Biology of the Cell

Article Title: SUMO-targeted ubiquitin ligase (STUbL) Slx5 regulates proteolysis of centromeric histone H3 variant Cse4 and prevents its mislocalization to euchromatin

doi: 10.1091/mbc.E15-12-0827

Figure Lengend Snippet: Slx5 prevents mislocalization of Cse4 to euchromatin. (A) Cse4 expressed from its own promoter is increased in chromatin fraction in psh1 ∆, slx5 ∆, and psh1 ∆ slx5 ∆ strains. Whole-cell extracts (WCEs) prepared from equal numbers of logarithmically growing cells in YPD were fractionated into soluble and chromatin fraction and assayed by Western blot analysis. Tub2 and histone H3 were used as markers for soluble and chromatin fractions, respectively. Two blots shown for Cse4 are indicated (L, long exposure; S, short exposure). (B, C) Cse4 expressed from its own promoter is mislocalized in psh1 ∆, slx5 ∆, and psh1 ∆ slx5 ∆ strains. Chromosome spreads were done by logarithmically growing cells in YPD. DAPI (blue) and α-HA (red) staining were used to visualize DNA and Cse4 localization, respectively. In wild-type strains, Cse4 is predominantly localized to one or two kinetochore clusters. In mutant strains, mislocalization of Cse4 is observed as multiple foci or diffused localization throughout the nucleus. The graph quantifies the number of cells exhibiting Cse4 mislocalization, and error bars are average deviation of two independent experiments. The number of cells used is indicated ( n ). Isogenic yeast strains used are wild type (YMB7290), psh1 ∆ (YMB7393), slx5 ∆ (YMB7588), and psh1 ∆ slx5 ∆ (YMB7607).

Article Snippet: Cells were visualized by DAPI staining (1 μg/ml in phosphate-buffered saline) mounted in antifade mountant (P36935; Molecular Probes, Eugene, OR).

Techniques: Western Blot, Staining, Mutagenesis

Journal: Cell reports

Article Title: Prdm12 Directs Nociceptive Sensory Neuron Development by Regulating the Expression of the NGF Receptor TrkA.

doi: 10.1016/j.celrep.2019.02.097

Figure Lengend Snippet: Figure 1. Prdm12 Expression in Dorsal Root Ganglia (A–W) Transverse DRG sections of embryonic or adult mice as indicated, double immunostained with Prdm12 (purple) and the indicated markers (green). (A) Immunostainings of Prdm12 and the neural crest transcription factor Pax3 at E11. (B–F) Immunostainings of Prdm12 and the pan- neuronal transcription factor Islet1 at E9.5 (B), E10.5 (C), E12.5 (D), or E14.5 (E). The proportion of Prdm12+, Islet1+, or Prdm12+/Islet1+ cells de- tected at the different analyzed stages is shown in (F) (mean ± SD; n = 2 [E10.5], 3 [E9.5, E12.5, and E14.5], or 5 [E11.5]) embryos. (G) Immunostaining of Prdm12 and a Sox10 reporter construct enabling the vizualisation of Sox10+ sat- ellite cells at E14.5. The inset shows a higher magnification view of an area with DAPI counter- staining (blue), and the arrowhead points to a putative Sox10+ satellite cell surrounding a Prdm12+

Article Snippet: Slides were washed with Tris-HCL buffer and mounted using Fluromount-G DAPI (Invitrogen).

Techniques: Expressing, Immunostaining, Construct, Staining