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Fig. 1 | Cell Communication and Signaling

Fig. 1

From: Clathrin- and dynamin-dependent endocytosis limits canonical NF-κB signaling triggered by lymphotoxin β receptor

Fig. 1

LTβR is internalized and trafficked towards degradation upon ligand binding. A549 cells were stimulated with Ago for the indicated time periods and immunostained for LTβR, EEA1 and LAMP1 (a) or trans- (TGN46) and cis-Golgi (GM130) (b). Insets show magnified views of boxed regions in the main images. Scale bars, 20 μm. Graphs represent the analysis of colocalization between LTβR and EEA1 or LAMP1, and integral intensity of LTβR (a) and colocalization between LTβR and GM130 or TGN46 (b). Data represent the means ± SEM, n ≥ 5 (a), n = 3 (b); ns - P > 0.05; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001 by one sample t test. c Lysates of A549 cells stimulated with Ago for different time periods were analyzed by Western blotting with antibodies against LTβR and vinculin (used as a loading control). Representative blots are shown. Values below blots represent the averaged LTβR/vinculin ratio (n = 5) in cells stimulated with Ago for the indicated time periods. Values are normalized to unstimulated control (time 0) set as 1.d Lysates of A549, HEK293T, CCD1070Sk and HeLa cells pretreated or not for 20 h (A549, HEK293T, CCD1070Sk) or 16 h (HeLa) with lysosomal degradation inhibitor, chloroquine (CQ), stimulated or not with Ago for the next 4 h were analyzed by Western blotting with antibodies against LTβR and GAPDH (used as a loading control). Representative blots are shown. Table presents the fold change of LTβR abundance in stimulated vs unstimulated cells (means, n ≥ 3)

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