Smarca2 inhibition accelerates the timeline of chromatin compaction. a Western blot for H3K9me3 (top), and α-tubulin (bottom) in embryos that were either mock injected or injected with 100 μM of the Smarca2 inhibitor PFI-3 at the one-cell stage. Protein was collected for analysis at 3.5 h post fertilization (hpf). b Western blot using embryos injected with increasing concentrations of PFI-3. Protein was collected for analysis at 3.5 hpf. c Electron micrographs demonstrating increased levels of chromatin compaction in 4.5 hpf embryos that were injected with dimethyl sulfoxide (DMSO) or 100 μM PFI-3 at the one-cell stage. Bottom panels represent higher magnification images (×20,000) of nuclear interior in mock- and PFI-3-injected embryos at specified time points. All scale bars indicate 1 μm. d Quantification of the number of particles per nuclear μm2 and percent nuclear area. Each data point indicates an individual embryo, for each embryo (four DMSO and nine PFI-3-injected embryos) values for particles per um2 and percent nuclear area were averaged from five to ten representitive nuclei. P values were calculated using the Student’s t test; error bars indicate SD. Source data for panels a and b is provided as a source data files
|
