believe the only way to fix the TSA is to end it, once and for all.
GSD super-resolution images of CaV1.2 channels in fixed ventricular myocytes or CaV1.2 channels and mCherry-Sec61β in fixed tsA-201 cells were generated using a Leica SR GSD 3D system.
Doses were fixed based on the time course (TSA 300 nℳ, 5-aza-CdR 10 μℳ and cisplatin 1 μℳ), and cytotoxicity at different time points (24 96 h) was measured to determine the optimal incubation time.
ES cell lines expressing FLAG (control), FLAG-Nup98FG, or FLAG-Nup98-HoxA9 either cultured in the absence or presence of TSA (50 nM, 24 hr) were fixed and stained with antibodies against FLAG (polyclonal) and indicated histone modifications.
The cells (4 × 10), seeded on sterile coverslips placed in 24-multiwell plates, were starved with or without 2.5 μ M TSA, treated with HGF (200 ng ml−1), fixed with 4% paraformaldehyde solution and permeabilised with 0.2% Triton X-100.
Although TSA is highly skewed, log10 (TSA) is approximately normally distributed; therefore, log10 (TSA) was fitted with a linear mixed model with (1) fixed effects for larva type (control or MO-injected fish) and time after injury (1, 2, 3, or 4 minutes) and (2) random effects for experimental day and larva within experimental day.
In addition, in Fig. 5, we compared the execution time of TSA to ESM by varying k with n fixed on random Gaussian matrices.
To determine whether HDAC activity is required for correct expression of Hdac1-regulated genes specifically during early neurogenesis, wild-type embryos were incubated with the HDAC inhibitor Trichostatin A (TSA) between 10 hpf and 14 hpf, then fixed and analysed for gene expression by in situ hybridisation.
Trichostatin A (TSA) was dissolved in DMSO to 3 μM and added to E3 medium to a final concentration of 1 μM. 10 hpf wild-type embryos were incubated in E3 medium containing 1 mM TSA for four hours until 14 hpf, then samples were fixed and analysed by in situ hybridisation Whole-mount in situ hybridisation was performed using standard procedures.
tsA-201 cells expressing CaV1.2 channels tagged at their C-terminus with monomeric GFP were fixed in 4% paraformaldehyde (10 min) and imaged in TIRF mode on the Leica 3D-GSD system described above using a 160×/1.43 NA objective.
