The pmCherry-Gal3 construct was a gift from Prof. Hemmo Meyer (Addgene plasmid # 85662; http://n2t.net/addgene:85662; RRID: Addgene_85,662). Raw264.7 cells (2.0 × 106) were transfected with 2 μg of pmCherry-Gal3 plasmid DNA using Nucleofector2b (Kit V, protocol D-032; Lonza) and seeded on 6-well plates. Twenty-four hours after transfection, cells expressing Gal3 with an N-terminal mCherry-tag were selectively grown in complete medium supplemented with 1000 μg/mL of G418 (Life Technologies) for 5 d. Monoclonal cell lines were further obtained by limited dilution in 96-well plates. Only clones with positive Gal3 puncta signals with LLoMe stimulation observed by fluorescent microscopy were used for the subsequent study. Cells stimulated either with LLoMe (2 mM) or with silica particles were fixed with 4% formaldehyde after 6 h and images were captured by fluorescent microscopy. The representative images from at least two independent experiments with biologically duplicated were shown in the result.

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