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. 2006 Oct 9;26(24):9232–9243. doi: 10.1128/MCB.01312-06

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FIG. 3. — Regulation of Akt activation by cell confluence. (A) Quiescent confluent (C) and nonconfluent (NC) mIMCD-3 cells plated for 24 h were stimulated in the presence or absence of HGF (40 ng/ml) for 10 min and immunoblotted with anti (α)-pAkt (upper panel) and α-Akt (lower panel). (B) Densitometric quantification of four independent experiments was performed as described for panel A. , P < 0.01 versus nonconfluent cells plus HGF. (C) Quiescent nonconfluent and confluent cells were treated with HGF for the indicated times (in minutes), and cell lysates were immunoblotted with α-pAkt (upper panel) and α-Akt (lower panel). (D) Quiescent nonconfluent cells were pretreated for 20 min in the presence or absence of 10 mM LY294002, then stimulated in the presence or absence of HGF for 10 min, and immunoblotted with α-pAkt (upper panel) and α-Akt (lower panel). (E) Quiescent confluent and nonconfluent mIMCD-3 cells plated for 24 h were stimulated in the presence or absence of HGF (40 ng/ml) for 10 min and immunoblotted with α-pERK (upper panel) and α-ERK (lower panel). (F) Densitometric quantification of four independent experiments was performed as described for panel E.

Inline graphic — Regulation of Akt activation by cell confluence. (A) Quiescent confluent (C) and nonconfluent (NC) mIMCD-3 cells plated for 24 h were stimulated in the presence or absence of HGF (40 ng/ml) for 10 min and immunoblotted with anti (α)-pAkt (upper panel) and α-Akt (lower panel). (B) Densitometric quantification of four independent experiments was performed as described for panel A. , P < 0.01 versus nonconfluent cells plus HGF. (C) Quiescent nonconfluent and confluent cells were treated with HGF for the indicated times (in minutes), and cell lysates were immunoblotted with α-pAkt (upper panel) and α-Akt (lower panel). (D) Quiescent nonconfluent cells were pretreated for 20 min in the presence or absence of 10 mM LY294002, then stimulated in the presence or absence of HGF for 10 min, and immunoblotted with α-pAkt (upper panel) and α-Akt (lower panel). (E) Quiescent confluent and nonconfluent mIMCD-3 cells plated for 24 h were stimulated in the presence or absence of HGF (40 ng/ml) for 10 min and immunoblotted with α-pERK (upper panel) and α-ERK (lower panel). (F) Densitometric quantification of four independent experiments was performed as described for panel E.