Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Jan 1;31(1):59–79. doi: 10.1091/mbc.E18-07-0469

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2020 Topalidou, Cattin-Ortolá, et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.

PMC Copyright notice

FIGURE 2: — Insulin localization is disrupted in Eipr1 KO cells. (A) Representative images of 832/13 (WT), Eipr1 KO 832/13 (Eipr1 KO), and Eipr1 KO 832/13 cells expressing WT Eipr1 (EIPR1[+]) costained for endogenous insulin and TGN38. In WT and EIPR1(+) cells, insulin is spread throughout the cytoplasm, but in Eipr1 KO cells insulin accumulates in a perinuclear region that partially overlaps with TGN38. The experiment was repeated three times, and the experimenter was blinded to the genotypes of the stained cells. Scale bars = 5 μm. (B) Pearson’s correlation coefficient was measured to quantify the localization between insulin and the TGN marker TGN38. n = 11 for WT, n = 13 for Eipr1 KO, and n = 15 for EIPR1(+); ***, p < 0.001; *, p < 0.05; ns, p > 0.05; error bars = SEM. The experiment was repeated three times. (C) Eipr1 KO cells have an increased Golgi/cytoplasmic ratio of insulin relative to WT cells. Fluorescence of a region of interest that includes the TGN divided by the fluorescence of a region of the same size in the cytoplasm, in WT and Eipr1 KO. (n = 25 for WT and Eipr1 KO; error bars = SEM; ***, p < 0.001). The data shown for the WT are the same as shown in Figure 2C of Cattin-Ortolá, Topalidou, et al. (2019) because these experiments were run in parallel with the same WT control. (D) Eipr1 KO cells have a slightly increased amount of insulin localized at or near the TGN. Fluorescence of a region of interest that includes the TGN divided by the fluorescence of a region of the same size in the background, in WT and Eipr1 KO. (n = 20 for WT and n = 30 for Eipr1 KO; error bars = SEM; *, p < 0.05). (E) Eipr1 KO cells have a decreased amount of insulin localized to the cytoplasm. Fluorescence of a region of interest in the cytoplasm divided by the fluorescence of a region of the same size in the background (n = 28 for WT and Eipr1 KO; error bars = SEM;***, p < 0.001). (F) Representative images of 832/13 (WT) and Eipr1 KO 832/13 (Eipr1 KO) cells costained for endogenous proinsulin and TGN38. In both WT and Eipr1 KO cells, proinsulin is localized in a perinuclear region that partially overlaps with TGN38. Scale bars = 5 μm. (G) Pearson’s correlation coefficient was measured to quantify the localization between proinsulin and the TGN marker TGN38. n = 18 for WT and n = 17 for Eipr1 KO; *, p < 0.05; error bars = SEM. The experiment was repeated three times.