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. 2019 Feb 1;10:22. doi: 10.3389/fendo.2019.00022

Figure 3.

Figure 3

The mature TCAP-1 peptide interacts with the HBD and a partial GAIN domain of LPHN1. (HBD constructs) LPHN1 HBD constructs were successfully expressed in HEK293 cells. (Inputs) Input lanes indicate strong presence of GFP-pro-mTCAP-1 or mature TCAP-1 in cell lysates prior to immunoprecipitation with HBD constructs V444-Q579 and V444-E634. Expected band size of GFP-pro-mTCAP-1 and GFP-mTCAP-1 are 40 and 30 kDa, respectively (black arrows). (IPs) Immunoprecipitation lanes show western blot resolution of corresponding eluates from input lanes. “No Ab” indicates that no anti-FLAG antibody was used to for immunoprecipitation of the HBD construct, serving as a negative control. No bands corresponding to GFP-pro-mTCAP-1 were isolated with either of the HBD constructs used for IP. A faint band at ~30 kDa, corresponding to GFP-mTCAP-1, is present when HBD V444-Q579 was used for IP. A stronger band of the same size is seen when IP was performed using HBD V444-E634 (red arrow). No bands corresponding to the GFP-pro-mTCAP-1 peptide were observed in the IP.