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Figure 5 | Cell Communication and Signaling

Figure 5

From: Intracellular signaling prevents effective blockade of oncogenic gp130 mutants by neutralizing antibodies

Figure 5

Effect of dominant-negative dynamin on surface expression and signaling of CAgp130. (A) and (B) T-REx-293 cells were transiently transfected with increasing amounts of an expression vector encoding dominant-negative K44A dynamin and GFP. (A) TCLs were analyzed by immunoblotting using Abs against dynamin, GFP and actin as loading control. (B) Cells were incubated with Alexa647 labeled transferrin. K44A dynamin expression and transferrin uptake were assessed via FACS analysis. (C) and (D) T-REx-293-CAgp130-mCherry were transfected with increasing amounts of dominant-negative K44A dynamin. Cells were left untreated or expression of CAgp130 was induced with 20 ng/ml dox for 24 h. (C) Overall receptor expression was assessed by FACS analysis of the fluorescent tag (right panel) and surface receptor expression was verified using the gp130 Ab B-P8 and an Alexa405 labeled secondary Ab (left panel). (D) TCLs were analyzed by immunoblotting using Abs against pStat3(Y705), dynamin, gp130 and actin as loading control.

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