Raffinose-Induced Lipid Accumulation in NHEK Cells

NHEK cells were seeded in 60-mm dishes and incubated overnight. After treatment with raffinose, the cells were washed twice with phosphate-buffered saline. The culture medium was changed every 48 h for raffinose treatment. Nile-red staining including fluorescence microscopy was performed as previously described39 (link). Immunocytochemistry was performed essentially as previously described using specific antibodies against involucrin (I9018, Sigma-Aldrich), loricrin (sc-51130, Santa Cruz Biotechnology), filaggrin (sc-66192, Santa Cruz Biotechnology), AQP3 (ab125219, Abcam), and LXR (PP-PPZ0412-10, Perseus Proteomics, Tokyo, Japan)39 (link).

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Na T.Y., Kim G.H., Oh H.J., Lee M.H., Han Y.H., Kim K.T., Kim J.S., Kim D.D, & Lee M.O. (2017). The trisaccharide raffinose modulates epidermal differentiation through activation of liver X receptor. Scientific Reports, 7, 43823.

Publication 2017

I9018 sc-51130 sc-66192 ab125219

After treatment Antibodies Cells Culture medium Dishes Filaggrin Fluorescence microscopy Immunocytochemistry Involucrin Loricrin Phosphate Raffinose Saline

Corresponding Organization :

Other organizations : Seoul National University

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Variable analysis

independent variables

Raffinose treatment

dependent variables

Lipid content (Nile-red staining)
Expression of differentiation markers (involucrin, loricrin, filaggrin, AQP3, LXR)

control variables

NHEK cells
Culture conditions (60-mm dishes, incubated overnight)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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