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. 2019 Feb;139(2):369–379. doi: 10.1016/j.jid.2018.08.020

Figure 5.

Figure 5

EGFR–kindlin-1 binding in human keratinocytes is EGF regulated. (a) SIM imaging of EGFR-GFP (green) and mCherry–kindlin-1 (magenta) after EGF stimulation. Inset boxes shown below each time point. (b) Immunoprecipitation of EGFR from KS cells re-expressing mCherry–kindlin-1 under starved conditions or after EGF stimulation (10 ng/ml). Graph on right shows quantification of five independent experiments. (c) Example lifetime images of KS cells expressing kindlin-GFP and mCherry–kindlin-1 after EGF stimulation. Graph on right shows quantification of 25 cells per condition over three experiments. (d) Example images of KS cells re-expressing mCherry–kindlin-1 (magenta) under growth conditions with DMSO or AG1478 treatment, fixed and stained for EGFR (green). Inset boxes shown below. Graph on right shows quantification of co-localization from 30 cells. (e) Immunoprecipitation of GFP or GFP–kindlin-1 under same conditions as in d, probed for phosphotyrosine (pY) and GFP. ∗P < 0.05, ∗∗P < 0.01 using two-way analysis of variance. Scale bars = 1 μm in a and 10 μm in c and d. EGFR, epidermal growth factor receptor; min, minute; SIM, structure illumination microscopy.