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. Author manuscript; available in PMC: 2017 Oct 2.
Published in final edited form as: Cell Rep. 2017 Sep 5;20(10):2480–2489. doi: 10.1016/j.celrep.2017.08.050

Figure 4. Granule cargo is retained in the trans-Golgi network following VGF knockdown.

Figure 4

832/3 cells (A, C, D) or GRINCH (B) cells were transfected with rat VGF siRNAs duplexes (SMARTpool) or a non-targeting control duplex (SCR). (A, B) Cells were treated with 2.5 mM Glc for 1 hr followed by 12 mM Glc for 30 minutes and lysates resolved on 8–20% iodixanol gradients. (A) Representative chromatograph of insulin-containing secretory granule fractions is shown (arrow indicates direction of increasing density). Solid line represents insulin; dotted line represents gradient density. (A, B) Immunoblots of gradient fractions are shown. (C) Immunostaining of insulin (green) relative to TGN46 (red) counterstained with DAPI (blue). (D) Immunostaining of CgB (green) relative to TGN46 (red) counterstained with DAPI (blue).