María Salazar, Arkaitz Carracedo, Íñigo J. Salanueva, Sonia Hernández-Tiedra, Mar Lorente, Ainara Egia, Patricia Vázquez, Cristina Blázquez, Sofía Torres, Stephane García, Jonathan Nowak, Gian María Fimia, Mauro Piacentini, Francesco Cecconi, Pier Paolo Pandolfi, Luis González-Feria, Juan L. Iovanna, Manuel Guzmán, Patricia Boya, Guillermo Velasco
ER stress precedes autophagy in cannabinoid action.
(A) Effect of THC on U87MG cell morphology. Note the presence of the
dilated ER in THC- but not vehicle-treated cells (6 h). Arrows point to the ER.
Scale bars: 500 nm. (B) Effect of SR1 (1 μM) and THC on
PDI immunostaining (red) in U87MG cells (8 h; n = 3). The
percentage of cells with PDI dots relative to the total cell number is shown in
the corner of each panel (mean ± SD). Scale bar: 20 μm.
(C) Effect of SR1 (1 μM) on THC-induced
eIF2α phosphorylation of U87MG cells (3 h; OD relative to
vehicle-treated cells, mean ± SD; n = 3).
(D) Effect of THC on PDI (red) and LC3 (green) immunostaining in
U87MG cells (n = 3). The percentage of cells with PDI or LC3 dots
relative to total cell number at each time point (mean ± SD) is shown.
Scale bar: 20 μm. (E) Effect of THC on eIF2α
phosphorylation and LC3 lipidation in U87MG cells (n = 3).
**P < 0.01 compared with THC-treated (B)
or vehicle-treated (C and D) cells.