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. 2020 May 6;295(25):8575–8588. doi: 10.1074/jbc.RA119.011979

Figure 1.

Figure 1.

Knockdown of endogenous tamalin inhibits the ligand-mediated endocytosis of mGluR1. A and B, Western blotting (A) and quantitation of the Western blots (B), showing the efficient knock-down of endogenous tamalin by shTam and replacement of the endogenous tamalin with full-length tamalin. C and D, Representative images (C) and quantitation of surface Myc-mGluR1 (D) showing that knockdown of endogenous tamalin with shTam and expression of the WT tamalin replacement construct had no effect on the surface expression of Myc-mGluR1 (n values: control, 35; shTam, 39; shTam:Tam, 38). E, Representative examples of surface and internalized Myc-mGluR1, 30 min after application of 100 μm R,S-DHPG in control cells, shTam-expressing cells, and shTam- and WT-tamalin-expressing cells. F, Quantitation of the endocytosis index suggested that knockdown of endogenous tamalin inhibited the R,S-DHPG-mediated internalization of Myc-mGluR1 and expression of WT tamalin rescued the normal trafficking of the receptor (n values: control, 35; control + DHPG, 36; shTam + DHPG, 38; shTam:Tam + DHPG, 39). The results are presented as means ± S.E. from three independent experiments. Scale bar, 10 μm. ***, p < 0.001; *, p < 0.05; n.s, p > 0.05.