Figure 3.
Raphin1 Inhibits R15B in Cells, Inducing a Transient Increase of eIF2α Phosphorylation and Attenuation of Protein Synthesis
(A, C, E, and G) Top: immunoblots of the indicated proteins in HeLa (A and C), R15a −/− (E), or R15b −/− (G) cells lysates treated with the indicated compounds at 10 μM for the indicated time. Bottom: quantifications of eIF2α phosphorylation in immunoblots as shown above. Data are means ± SEM; n = 3. ∗p < 0.05; ∗∗p < 0.01 by an unpaired two-tailed Student’s t test in comparison to 0 hr time point. ns, not significant.
(B, D, F, and H) Upper panel: autoradiogram of newly synthesized proteins radiolabeled with 35S-methionine in HeLa (B and D), R15a −/− (F), or R15b −/− (H) cells treated with the indicated compounds at 10 μM for the indicated time. Lower panel: Coomassie-stained gel.
Representative results of three independent experiments are shown.
(I) Cartoon illustrating the activity of Raphin1.
See also Figures S3 and S4.