![TPRN interaction with actin and oligomerization. (A) Diagram of the TPRN constructs used for biochemical experiments. (B and C) TPRN interacts with β-actin. HEK293 cells were transfected with the constructs indicated for each panel. Immunoprecipitations were carried out with Myc antibody, followed by western blotting to detect co-expressed proteins. The upper rows show Co-IP results, and the lower rows show input protein. IP: Myc shows IP of all the constructs with Myc tag. (B) HA–β-actin is pulled down by FL Myc–TPRN, indicating its interaction with TPRN. No HA–β-actin was pulled down by Myc antibody without TPRN. (C) Each contiguous region of TPRN, (1–170, 171–410, 411–622, and 623–749 [RefSeq: NP_780495.2]) can mediate interactions with F-actin. Please see additional results in Fig. S1, J–L. (D) Co-IP shows homomeric interactions between FL TPRN–EGFP and Myc–TPRN. (E–J) Homomeric interactions of TPRN fragments illustrated by Co-IP. TPRN1–410-EGFP and Myc-TPRN1–410, TPRN411–749-EGFP and Myc-TPRN411–749, TPRN171–410-EGFP and Myc-TPRN171–410, TPRN411–622-EGFP and Myc-TPRN411–622, TPRN623–749-EGFP and Myc-TPRN623–749, and HA-TPRN1–170 and Myc-TPRN1–170 can oligomerize. Molecular weight markers (kDa) are shown on the left side of each blot. Source data are available for this figure: SourceData F4.](https://cdn.rupress.org/rup/content_public/journal/jcb/224/8/10.1083_jcb.202408026/2/jcb_202408026_fig4.png?Expires=1771584471&Signature=kq6gSqkxRpVAecPcH~rqA5kaaWqweUl6T9sA0S0HUZKQpaeD2Iu5mR7yRNMVm4xGatrft4PV53cVJqJtsD7guLfL6B-AbZXMIEwFh5DV9AUq531tSj5dZioMwTXU3Btz0FTI1vD8nlOGIzHbmk~z1Fc5U5kQYa1vngkwvrVgQMrumOVW6gZyyC1MOubbV8bbghkVSj0T7Brekn55eH4COL7qAoKpRP4tkTEPoyg0G0ylkq8jaqYdrX20muxELdUGRlQdsG031-ZOGJKM8ZnoAjK~tdCDwxuijx1nb8wKHwgtk04dFxjentAR2CAqaM214b1TPW8J82~vtZpp4Hmh1Q__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
TPRN interaction with actin and oligomerization. (A) Diagram of the TPRN constructs used for biochemical experiments. (B and C) TPRN interacts with β-actin. HEK293 cells were transfected with the constructs indicated for each panel. Immunoprecipitations were carried out with Myc antibody, followed by western blotting to detect co-expressed proteins. The upper rows show Co-IP results, and the lower rows show input protein. IP: Myc shows IP of all the constructs with Myc tag. (B) HA–β-actin is pulled down by FL Myc–TPRN, indicating its interaction with TPRN. No HA–β-actin was pulled down by Myc antibody without TPRN. (C) Each contiguous region of TPRN, (1–170, 171–410, 411–622, and 623–749 [RefSeq: NP_780495.2]) can mediate interactions with F-actin. Please see additional results in Fig. S1, J–L. (D) Co-IP shows homomeric interactions between FL TPRN–EGFP and Myc–TPRN. (E–J) Homomeric interactions of TPRN fragments illustrated by Co-IP. TPRN1–410-EGFP and Myc-TPRN1–410, TPRN411–749-EGFP and Myc-TPRN411–749, TPRN171–410-EGFP and Myc-TPRN171–410, TPRN411–622-EGFP and Myc-TPRN411–622, TPRN623–749-EGFP and Myc-TPRN623–749, and HA-TPRN1–170 and Myc-TPRN1–170 can oligomerize. Molecular weight markers (kDa) are shown on the left side of each blot. Source data are available for this figure: SourceData F4.
