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. 2017 Apr 10;7:46064. doi: 10.1038/srep46064

Figure 1. Stable cell lines used in this study.

Figure 1

(a) RBL-2H3 cells were stably transfected with pApuro-myc-His-Mincle WT or pApuro-myc-His-Mincle R42I mutant by electroporation. Clones resistant to puromycin were selected and screened for the level of protein expression. Two cloned cell lines of each transfectant were solubilized in lysis buffer. Precleared lysates were analyzed by immunoblotting with anti-myc and anti-FcεRIβ mAbs, respectively. (b) Analysis of cell surface expression of Mincle by flow cytometry. Cells were stained with an Alexa Fluor 488-labeled anti-myc mAb (solid line) or Alexa Fluor 488-labeled control mouse IgG1 (dashed line). Data are representative of three independent experiments. (c) Detergent-soluble lysates (input) and anti-myc immunoprecipitates (IP) from RBL-2H3 cells and cells expressing WT Mincle (PA-11, WT) or the R42I mutant (R42I-3, R42I) were analyzed by immunoblotting with the indicated antibodies. Similar results were obtained from the other cloned cell lines. (a and c) Molecular size markers are indicated at the left in kDa. Data are representative of three independent experiments.