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

. 1999 Jun;19(6):4008–4018. doi: 10.1128/mcb.19.6.4008

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

Copyright © 1999, American Society for Microbiology

PMC Copyright notice

FIG. 3 — Transient transfection of Δp85α PI 3-kinase inhibits, and GagPKB potentiates, insulin-stimulated translocation of GLUT4myc in muscle cells. L6-GLUT4myc myoblasts were transiently transfected with GFP (0.4 μg) (A) and cotransfected with HA-PKB (0.4 μg) (B), dominant-negative Δp85α (0.4 μg) (C), or GagPKB (0.4 μg) (D) and incubated for 48 h in culture. (A) Cells were left untreated (basal) or treated with 100 nM insulin for 20 min (insulin) and then processed for cell surface GLUT4myc detection with anti-myc antibody (1:100) followed by Cy3-conjugated goat anti-mouse antibody as indicated in Materials and Methods. Each pair of panels (upper and lower) shows the same field of cells. In the lower panels, GFP fluorescence in transfected cells (arrowheads) is shown. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of the transfected cells. Results shown are representative of at least five experiments. (B) Cells were fixed and processed for indirect immunofluorescence with anti-HA antibody (α-HA; 1:1,000) followed by detection with a Cy3-conjugated goat anti-mouse antibody. The panel on the left shows the number of cells in the field seen by phase-contrast microscopy. The middle and right panels show the GFP-positive cells and the HA-PKB-positive cells, respectively, in the same field. The results indicate complete overlap of GFP- and HA-PKB-positive cells. Results shown are representative of three experiments. (C) Cells were treated as described for panel A. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of cells transfected with Δp85α. Results shown are representative of three experiments. (D) Cells were treated as described for panel A. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of cells transfected with GagPKB. Results shown are representative of three experiments.

FIG. 3 — Transient transfection of Δp85α PI 3-kinase inhibits, and GagPKB potentiates, insulin-stimulated translocation of GLUT4myc in muscle cells. L6-GLUT4myc myoblasts were transiently transfected with GFP (0.4 μg) (A) and cotransfected with HA-PKB (0.4 μg) (B), dominant-negative Δp85α (0.4 μg) (C), or GagPKB (0.4 μg) (D) and incubated for 48 h in culture. (A) Cells were left untreated (basal) or treated with 100 nM insulin for 20 min (insulin) and then processed for cell surface GLUT4myc detection with anti-myc antibody (1:100) followed by Cy3-conjugated goat anti-mouse antibody as indicated in Materials and Methods. Each pair of panels (upper and lower) shows the same field of cells. In the lower panels, GFP fluorescence in transfected cells (arrowheads) is shown. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of the transfected cells. Results shown are representative of at least five experiments. (B) Cells were fixed and processed for indirect immunofluorescence with anti-HA antibody (α-HA; 1:1,000) followed by detection with a Cy3-conjugated goat anti-mouse antibody. The panel on the left shows the number of cells in the field seen by phase-contrast microscopy. The middle and right panels show the GFP-positive cells and the HA-PKB-positive cells, respectively, in the same field. The results indicate complete overlap of GFP- and HA-PKB-positive cells. Results shown are representative of three experiments. (C) Cells were treated as described for panel A. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of cells transfected with Δp85α. Results shown are representative of three experiments. (D) Cells were treated as described for panel A. The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b). Arrowheads indicate the positions of cells transfected with GagPKB. Results shown are representative of three experiments.