Lastly, mTORC1 contributes to tumor cell growth by inhibiting autophagy, a catabolic pathway that degrades aged or damaged organelles (Figure 1(d)); mTORC1 suppresses autophagy directly by inhibiting the kinase complex unc-51-like kinase 1/mammalian autophagy-related gene 13/focal adhesion kinase family-interacting protein of 200 kDa (ULK1/Atg13/FIP200) which is a key component required for the autophagy induction and indirectly by modulating the expression of death-associated protein 1 (DAP1
), a novel substrate of mTORC1 that negatively regulates autophagy [13, 14] (Figure 1(c)).
Immunofluorescence results revealed decreased [beta]-catenin immunofluorescence (green) in the cells treated with EGCG; DAP1
counterstaining (blue) was similar in the cells treated with or not treated with EGCG (Fig.
Blocked eggs were then incubated >2 h to overnight in primary antibodies, followed by a 2-h incubation period in the appropriate fluorophore-conjugated, secondary antibodies, then mounted in ProLong antifade reagent containing 4',6-diamidino-2-phenylindole (DAP1
; Thermo Scientific) in order to stain chromosomes.
The sperm was then stained (on the filter) for 10 minutes with 4',6diamidino-2-phenylindole (DAP1
) flourescent nuclear stain.
Panel (a) overlay of HBcAg (red) and EpCAM (green) positive cells and DAP1
nuclei counterstaining (blue) in HCC tissue (panels (a1)-(a3), resp.).
as a fluorescence DNA binding agent was applied to assess nuclear apoptosis.
Foram selecionadas oito arvores com os respectivos diametros iguais ou proximos da media entre as trinta arvores, e avaliadas a altura total, a altura da copa, o diametro a altura do peito (DAP1
,30 m) com o uso de trema e o peso da copa, considerando folhas e galhos, com uso de uma balanca de pesagem suspensa.
mTORC1 regulates WIPI2 (mammalian orthologue of Atg18), which is also essential for phagophore formation , as well as DAP1
(death associated protein 1), an inhibitor of autophagy .
. GFP, phase contrast, and bright field images are overlaid with some artistic license."
Immunostained using mouse anti-Src antibody (dilution in 1:100) and Alexa Fluor 488 conjugated goat anti-mouse IgG (1:1000; ThermoFisher Scientific), and counterstained for nuclei with DAP1
. Immunofluorescence was visualized using a confocal laser scanning microscope (Carl Zeiss Inc.).