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Fig. 3 | Cell Communication and Signaling

Fig. 3

From: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling

Fig. 3

Cholesterol replenishment rescues the effects of MβCD treatment on LTβR signaling and internalization. a Schematic description of cholesterol replenishment experiments. b Lysates of A549 cells treated as depicted in a were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control. Graphs depict densitometric analysis for the indicated proteins from Western blotting (protein levels normalized to vinculin). Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n = 5; ns - P > 0.05; *P ≤ 0.05; ***P ≤ 0.001 by ANOVA test. c Immunofluorescence staining of ligand-bound LTβR and EEA1 in A549 cells treated as depicted in A except Step 4, where cells were fixed and stained instead of cell lysis. Insets: magnified views of boxed regions in the main images. Scale bars, 20 μm. d Analysis of integral intensity and the number of LTβR- and EEA1-positive vesicles in cells treated as in C. Values are presented as fold change versus controls - vehicle-treated cells marked as a black line, set as 1. Data represent the means ± SEM, n = 3. ns - P > 0.05; *P ≤ 0.05; **P ≤ 0.01 by one sample t-test

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