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. 2016 May 12;15(8):1648–1659. doi: 10.1016/j.celrep.2016.04.062

Figure 1.

Figure 1

Validation of Depletion of TFG using siRNA

(A) Immunoblotting of HeLa cell lysates from cells transfected with siRNAs targeting Lamin A/C or TFG or with a non-targeting control (GL2) as indicated. Four independent duplexes were used against TFG. Blots were probed to detect TFG, Sec16A, Lamin A/C, or α-tubulin as indicated.

(B and C) Immunoblotting of (B) RPE-1 or (C) IMR-90 cell lysates from cells transfected with siRNAs targeting GL2 or TFG as indicated showing effective depletion of TFG compared to the dynein intermediate chain subunit DIC74.

(D–I) Early secretory pathway organization in TFG-depleted cells. Widefield immunofluorescence microscopy of control or TFG-depleted cells labeled to detect (D) Sec16A and GRASP65, (F) Sec24C and GM130, and (H) Sec31A and GalT. RGB merges include DAPI labeling of nuclei. Green-magenta merges are also shown. (D), (F), and (H) show automated quantification of puncta numbers per μm2 for (E) Sec16A, (G) Sec24C, and (I) Sec31A.