3i, j). Expectedly, the frequency and the number of pre-HSC IIminus (pre-HSC II without pre-HSC IIplus, representing mature hematopoietic cells) was decreased more dramatically (Supplementary Fig. 3o), demonstrating that Atg5 deletion promotes pre-HSC emergence and blocks their maturation. ...
We used Maxima H Minus cDNA synthesis Master Mix (ThermoFisher Scientific, TERM1662) to generate cDNAs from RNAs and SYBR Green Master Mix (ThermoFisher Scientific, A46113) for qPCR reaction following the manufacturer’s protocols. Total cDNA of 10 ng was used for each reaction in 96-well...
As quantified by western blots, autophagic flux was defined as the quantity of LC3b-II or p62 (expressed as ng, pmol, or normalized values) in the leupeptin treated sample minus the average quantity in the sham treated samples (Haspel et al., 2011). Although other groups adopt the practice...
Therefore, it can mediate the plus-end and minus-end transport of vesicles. The transport of autophagosomes and lysosomes toward each other is essential for fusion. Once the autophagosomes and lysosomes reach their destination, they tether to their partner organelle. Large coiled-coil proteins or ...
Band intensity was determined by densitometry with NIH Image J software (v1.48h3) using mean gray value minus background. In some experiments, cells were treated with bafilomycin A1 (50 nM, #S1413, SellectChem) at 37 °C prior to lysis. 2.7. Immunoprecipitation For immunoprecipitation, ...
The number of RGCs was calculated by subtracting from the amount of γ-Synuclein positive cells minus the displaced amacrine double stained γ-Synuclein and Calbindin cells. The control eyes are represented as baseline value, n = 4 animals per group. #p < 0.01 in comparison to the ...
of SIN minus-strand RNA levels (s) and HSV-1 immediate early gene ICP27 mRNA levels (t) of triplicate samples at indicated time point, respectively. u, v, Quantitative real-time PCR analyses of relative mRNA levels of interferon beta (IFN-β; u) and the interferon-stimulated gene (ISG)...
The coordinates relative to bregma were, anteroposterior: −6.30 mm, −7.80 mm; lateral, ±1.50 mm; dorsoventral, −1.50 mm (minus indicates a position posterior to bregma). Correlative light and electron microscopy (CLEM) For the merging of confocal fluorescence microscopy ...
8A; compare lane 6 minus lane 5 versus lane 2 minus lane 1), statistical significance was not obtained (Fig. 8B). Similar experiments were performed in MEFs. Consistently, we observed that lack of endophilin B2 expression significantly suppressed starvation-induced autophagic flux (Fig. 8, C ...
Our research complies with all relevant ethical regulations and guidelines. All zebrafish procedures were performed in accordance with the UK Animals (Scientific Procedures) Act with appropriate Home Office Project and Personal animal licences and with local Ethics Committee approval. The studies were perf...