To examine histone modifications in differentiating germ cells, we stained wild-type ovaries using monoclonal antibodies specific for histone modifications (Fig. 1; Kimura et al., 2008).
In macronuclear anlagen antibodies directed against these modifications neither stained the banded regions nor the heterochromatic blocks of polytene chromosomes, and therefore these modifications are probably localized in the interbands of the polytene chromosomes, but cannot be detected on the level of resolution used here.
To investigate whether inhibition of cell death induced by iPTH is associated with bone matrix modifications, we quantified the prevalence of osteocytes death in cortical bone stained using the TUNEL assay.
To further explore whether loss of Spen affected Xist-mediated chromatin modifications, we performed Xist RNA fluorescence in situ hybridization (FISH) and immunofluorescence staining.
The sections were prepared as described above, with the following modifications: After staining for GATA-4 sc-12377) or GATA-6 sc-90555) with 3,3'-diaminobenzidine, the slides were incubated with the second primary antibody for 1 h at +37°C.
Except slight modifications, staining of primary hippocampal neurons was performed as described previously [37].
In general, all assays were performed according to the manufacturer's instructions with minor modifications to optimize staining.
The humiliation as we discover you stained the sheets.
To further characterise the distribution of the H4R3me2s and H4R3me2a modifications during neural development, I stained cortices from E15.5 with the antibodies against these histone modifications and an anti-NeuN antibody to identify post-mitotic neurons.
2D gels were either Coomassie-blue stained or stained using a Mass Spectrometry-compatible silver staining protocol that is based on a modification of the EMBL silver staining protocol [42].
