Figure S2.

G3BP1 inhibits mTORC1 in cells without SGs, related to Figure 1

(A) Insulin/aa-stimulated siG3BP1 cells. n = 6.

(B) Quantitation of G3BP1 in (A). Shown are data points and mean ± SEM.

(C) Quantitation of RPS6KB1-pT389 in (A). Data shown as in (B).

(D) Quantitation of RPS6-pS235/236 in (A). Data shown as in (B).

(E) Quantitation of G3BP1 in (G). Mean ± SEM.

(F) Quantitation of RPS6KB1-pT389 in (G). Data shown as in (E).

(G) Time course analysis of shG3BP1 #1 cells, insulin/aa-stimulated for up to 30 min. n = 3.

(H) shG3BP1 #1 cells cultured in full medium. n = 7.

(I) Quantitation of G3BP1 in (H). Shown are data points and mean ± SEM.

(J) Quantitation of RPS6KB1-pT389 in (H). Data shown as in (I).

(K) shG3BP1 #2 cells cultured in full medium. n = 4.

(L) Quantitation of G3BP1 in (K). Shown are data points and mean ± SEM.

(M) Quantitation of RPS6KB1-pT389 in (K). Data shown as in (L).

(N) Serum/aa-starved shG3BP1 #1 cells. Arrow, RPS6KB1-pT389 signal. n = 8, including re-analysis of improved contrast detections for data shown in Figures 1F–1H.

(O) Quantitation of G3BP1 in (N). Shown are data points and mean ± SEM. n = 8, including re-analysis of data shown in Figures 1F–1H.

(P) Quantitation of RPS6KB1-pT389 in (N). Data shown as in (O). n = 8, including re-analysis of improved contrast detections for data shown in Figures 1F–1H.

(Q) IF of shG3BP1 #1 cells. Cells were either serum/aa-starved and stimulated with insulin/aa for 15 min; or serum-starved and treated with arsenite for 30 min. Overlay: white, EIF3A and G3BP1 co-localization; magenta, EIF3A; green, G3BP1; inserts, magnifications of yellow square. Scale bar, 10 μm. n = 3, except shG3BP1 #1, arsenite [0 min], n = 2.

(R) Quantitation of data shown in (Q). Shown are data points and mean ± SEM.

(S) Immunoblot performed in parallel to IF data in (Q). Insulin/aa-stimulated shG3BP1 #1 cells. n = 3.

(T) Quantitation of G3BP1 in (S). Shown are data points and mean ± SEM.

(U) Quantitation of RPS6KB1-pT389 in (S). Data shown as in (T).

(V) Immunoblot performed in parallel to IF data in (Q). Arsenite-exposed shG3BP1 #1 cells. n = 3.

(W) Quantitation of G3BP1 in (V). Shown are data points and mean ± SEM.

(X) Quantitation of RPS6KB1-pT389 in (V). Data shown as in (W).