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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: Biochim Biophys Acta Mol Cell Biol Lipids. 2019 Jun 5;1864(10):1514–1524. doi: 10.1016/j.bbalip.2019.05.017

Figure 3. Scp2-like domain in Hsd17b4 interacts with ceramide.

Figure 3

A. Domain structure of Hsd17b4 and EGFP-tagged constructs used for transfection. B. HEK293T cells were transfected with EGFP-tagged constructs encoding the three domains of Hsd17b4 or full length protein as depicted in A. Expression products were isolated from cell lysates using affinity chromatography of on ceramide beads, followed by SDS-PAGE and immunoblotting with anti-GFP antibody. Expression and affinity chromatography of EGFP was used as the negative control. Lane 1, marker; lanes 2-6, input; lane 7-11, output (pulled-down protein). C. Immunocytochemistry using anti-ceramide antibody and EGFP-tagged Scp2-like domain from Hsd17b4 (hsdSCP2) ectopically expressed in HEK293T cells.