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Figure S4
G3BP1 phenocopies lysosomal TSC functions, related to
(A) PLA of G3BP1-TSC2 in serum/aa-starved siG3BP1 cells. PLA puncta, white dots; nuclei, blue (DAPI). Scale bar, 10 μm. n = 3.
(B) Quantitation of data in (A). Shown are data points and mean ± SEM. n = 5 technical replicates.
(C) Sucrose density gradient of serum/aa-starved or insulin/aa-stimulated MCF-7 cells. n = 3.
(D) Quantitation of data in (C). Area under the curve highlighted in green (G3BP1), orange (TSC2), and gray (LAMP2), starved condition; dashed lines, stimulated condition. Mean ± SEM.
(E) Insulin/aa-stimulated shG3BP1 #1 cells. n = 5.
(F) Quantitation of TSC2-pT1462 in (E). Shown are data points and mean ± SEM.
(G) Quantitation of AKT1-pS473 in (E). Data shown as in (F).
(H) Insulin/aa-stimulated siTSC2 cells. n = 6.
(I) Quantitation of TSC2 in (H). Shown are data points and mean ± SEM.
(J) Quantitation of RPS6KB1-pT389 in (H). Data shown as in (I).
(K) Cell size of TSC2 KO cells. Mean ± SEM. *p < 0.05. n = 3.
(L) IF of LAMP2 positioning in serum/aa-starved siG3BP1 cells. White, LAMP2; blue (Hoechst), nuclei. Scale bar, 10 μm. n = 3.
Reprinted from Cell, 184(3), Prentzell, M.T., Rehbein, U., Cadena Sandoval, M., De Meulemeester, A.S., Baumeister, R., Brohée, L., Berdel, B., Bockwoldt, M., Carroll, B., Chowdhury, S.R., von Deimling, A., Demetriades, C., Figlia, G., Genomics England Research Consortium, de Araujo, M.E.G., Heberle, A.M., Heiland, I., Holzwarth, B., Huber, L.A., Jaworski, J., Kedra, M., Kern, K., Kopach, A., Korolchuk, V.I., van 't Land-Kuper, I., Macias, M., Nellist, M., Palm, W., Pusch, S., Ramos Pittol, J.M., Reil, M., Reintjes, A., Reuter, F., Sampson, J.R., Scheldeman, C., Siekierska, A., Stefan, E., Teleman, A.A., Thomas, L.E., Torres-Quesada, O., Trump, S., West, H.D., de Witte, P., Woltering, S., Yordanov, T.E., Zmorzynska, J., Opitz, C.A., Thedieck, K., G3BPs tether the TSC complex to lysosomes and suppress mTORC1 signaling, 655-674.e27, Copyright (2021) with permission from Elsevier. Full text @ Cell