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. 2021 Feb 22;6(4):e143324. doi: 10.1172/jci.insight.143324

Figure 2. The I154T substitution disrupts the interaction of 2 ECDs within an mGlu7 dimer.

Figure 2

(A) Location of isoleucine 154 in the structure of the mGlu7 ECD. PBD 2E4Z. (B) Protein sequence alignment of all 8 human mGlu receptor subtypes. Darker shades of blue indicate increased conservation. (C) Western blot of total protein lysates and media from cells expressing HA-ECD-WT and HA-ECD-I154T constructs in the absence of DTT. (D) Western blot of total protein lysates and media from cells expressing HA-ECD-WT and HA-ECD-I154T constructs in the presence of DTT. (E) Experimental design for cell surface ELISA experiments. (F–H) Quantification of (F) intracellular HA signal, (G) surface HA signal, and (H) surface MYC signal. n = 3 independent experiments with 3 technical replicates each. One-way ANOVA with Tukey’s multiple comparisons. ****P < 0.0001. ns, not significant; ECD, extracellular domain; mGlu7, metabotropic glutamate receptor subtype 7; FL, full-length.