Figure 4.

G3BP1 tethers the TSC complex to lysosomes

(A) PLA of TSC2-LAMP2 in insulin/aa-stimulated siG3BP1 cells (15 min, 1 μM insulin). PLA puncta, white dots; nuclei, blue (DAPI). Scale bar, 100 μm. n = 4.

(B) Quantitation of data in (A). Shown are data points and mean ± SEM. Control (0 min) normalized to 1. n = 12 technical replicates.

(C) IF of LAMP1-TSC2 co-localization in G3BP1 KO cells transfected with siRHEB; insulin/aa stimulation (1 μM insulin). Overlay: white, LAMP1-TSC2 co-localization; green, TSC2; magenta, LAMP1; insert, magnification of the yellow square. Scale bar, 10 μm. n = 3.

(D) Quantitation of data in (C). Shown are data points and mean ± SEM.

(E) TSC2 KO cells in full medium. n = 3.

(F) Quantitation of TSC2 in (E). Shown are data points and mean ± SEM.

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

(H) Size of G3BP1 KO cells. Mean ± SEM. *p < 0.05. n = 3.

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

(J) IF of MTOR-LAMP2 co-localization in G3BP1 KO cells. Overlay: white, MTOR-LAMP2 co-localization; green, MTOR; magenta, LAMP2; insert: magnification of the yellow square. Scale bar, 10 μm. n = 3.

(K) Insulin-stimulated TSC2 KO cells transfected with siG3BP1. n = 4.

(L) Quantitation of TSC2 in (K). Shown are data points and mean ± SEM. TSC2 was compared between control and TSC2 KO cells.

(M) Quantitation of G3BP1 in (K). Shown are data points and mean ± SEM. G3BP1 was compared between siControl and siG3BP1 in control or TSC2 KO cells.

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

See also Figure S4.