Figure 6.

Insulin-induced integrin α3 and LN332 production contributes to keratinocyte migration. (A, B) Cells seeded in chamber slides were either left untreated or treated with 10-7 M insulin for 24 h, followed by immunolabeling with the anti-integrin α3 Ab and FITC-conjugated secondary Ab. (A), control (B), insulin treatment 24 h. (C) Non-reducing western blot analysis of integrin α3 expression shows stimulation of this integrin by insulin in keratinocytes. Results are representative of three independent experiments. Levels of integrin α3 were quantified by determining the ratio of the integrated density of integrin α3 to GAPDH loading control using ImageJ software. Data are shown as the mean value +/- SD. *P < 0.05, **P < 0.01 vs control treatment. (D, E) Cells seeded in chamber slides were either left untreated (D), or treated with 10-7 M insulin for 24 h (E), followed by immunolabeling with the anti-LN332 Ab. Small boxes show higher magnification for more detailed distribution of the staining. (F) Non-reducing immunoblot analysis of LN332 levels. Insulin stimulates secretion of LN332 by keratinocytes. Results are representative of three independent experiments. Levels of LN332 were quantified by determining the ratio of the integrated density LN332 to GAPDH loading control using ImageJ software. Data are shown as the mean value +/- SD. *P < 0.05 vs control treatment. (G) Cells were wounded by scratching and then pre-treated with 25 μg/ml of function-inhibiting Abs directed against LN332 (P3H9-2) and/or integrin α3 (P1B5) for 1 h, then treated with 10-7 M insulin. Migration distances were measured after 24 and 48 h of treatment. *P < 0.05 vs control or as indicated. Insulin induced keratinocyte migration was eliminated by both integrin α3 and LN332 function-inhibiting Abs. ***P < 0.001 vs control or as indicated. Each treatment group was performed in triplicate.

Liu et al. BMC Cell Biology 2009 10:1   doi:10.1186/1471-2121-10-1
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