Figure 2
From: Hierarchical scaffolding of an ERK1/2 activation pathway
IQGAP1 and KSR1 interact with ERK1/2 in a stimulus-specific manner. (A) Lysates of unstimulated, DPBA-stimulated or FCS-stimulated cells were subjected to immunoprecipitation (IP) experiments. Co-immunoprecipitated proteins were detected by western blotting and analyzed by densitometry. (A) The graph shows average band intensities of ERK1/2 co-immunoprecipitated with an anti-IQGAP1 antibody. (B) Representative western blot for an anti-IQGAP1 IP experiment. (C) Average band intensities of ERK1/2 co-immunoprecipitated with an anti-KSR1 antibody. (D) Representative western blot for an anti-KSR1 IP experiment. (E) Average band intensities of IQGAP1 co-immunoprecipitated with an anti-ERK1/2 antibody. (F) Average band intensities of KSR1 co-immunoprecipitated with an anti-ERK1/2 antibody. (G) Representative western blot for an anti-ERK1/2 IP experiment. Please note that a non-specific band of 38 kDa (*) was seen in ERK1/2 IPs. (H) Analysis of pERK to total ERK ratios in input samples as well as in immunoprecipitates. Significance (*compared to unstimulated samples; # compared to ) and p-values are indicated on chart; error bars represent standard errors. IP, immunoprecipitation. Please note that ERK2 appears as a doublet after stimulation, with the upper band representing phosphorylated ERK2.