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. 2017 Mar 19;292(18):7588–7597. doi: 10.1074/jbc.M117.785345

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 10. — CerS and ACBP interact in situ. FLAG-tagged ACBP was co-expressed in HEK293T cells together with HA-tagged CerS2 or CerS3 (A), the indicated HA-tagged CerS (B), and HA-tagged wild-type and mutated (both W15R) forms of CerS2 or CerS3 (C). PLA using primary antibodies against HA and FLAG tags and secondary antibodies carrying PLA probes were applied to visualize the ACBP-CerS interaction. DAPI was used to detect nuclei. Scale bars, 10 or 20 μm (ACBP/CerS2 in B). Controls were cells transfected with empty plasmids alone or in combination with either ACBP or CerS3 plasmids. Other controls received either only one primary antibody or none. D, Western blots show expression of FLAG-tagged ACBP and HA-tagged CerS in HEK293T cells transfected with the indicated plasmid. GAPDH was used as loading control. The arrows mark the CerS. 2_m and 3_m indicate cells expressing CerS2 W15R and CerS3 W15R, respectively.