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. 2018 May 25;293(28):11047–11066. doi: 10.1074/jbc.RA118.001747

Figure 1.

Figure 1.

LC1/BAD domain of U1-70K is necessary and sufficient for self-association. A, full-length (WT) recombinant GST-fused, Myc-tagged U1-70K (rU1-70K) and variants lacking one or both LC domains (ΔLC1, ΔLC2, and ΔLC1 + ΔLC2) were overexpressed in HEK293 cells and immunoprecipitated (IP) with anti-Myc antibodies. IP with a nonspecific IgG was also performed from mock-transfected cells as a negative control. Western blotting for recombinant Myc-tagged proteins (green) and native U1-70K (red) are shown for both the inputs and co-IPs (A, bottom panels). B, full-length WT and rU1-70K truncations, including the N terminus (1–99 residues) alone, the N terminus and RRM (1–181 residues), LC1/BAD alone (231–308 residues), and the LC2 domain alone (residues 317–407). IP with a nonspecific IgG was also performed from mock-transfected cells as a negative control. Western blotting for recombinant Myc-tagged proteins (green) and native U1-70K (red) are shown for both the inputs and co-IPs. C, WT rU1-70K was immunoprecipitated from untreated and RNase (50 ng/μl)-treated lysates followed by Western blotting for the Myc tag recombinant protein (green) and native U1-70K (red). IP with a nonspecific IgG was also performed from mock-transfected cells as a negative control.