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. 2023 Jan;29(1):97–110. doi: 10.1261/rna.079320.122

FIGURE 4.

FIGURE 4.

Alternative splicing factor TIA1 directly interacts with U1C and PRPF39. (A) Nam8 (pink) contacts U1C (blue) and Prp42 (yellow) in the yeast U1 snRNP structure (PDB 6N7R). N, C, and 61 indicate the amino- and carboxy-terminal ends and residue 61 (which separates the NTD and CTD) of U1C. (B) Purified GST-U1C FL and NTD (bottom, Coomassie stained SDS PAGE) can pull down TIA1 (top, western blot with anti-TIA1 antibody). (C) A schematic diagram of TIA1 domains and the three TIA1 truncation constructs used in subsequent experiments. (D) Purified GST-RRM1 and RRM3 + Q, but not RRM2 (bottom, Coomassie stained SDS PAGE), can pull down FL U1C (top, western blot with anti-U1C antibody). (E) GST pull-down experiment using GST-TIA1 as a bait (bottom, Coomassie stained SDS PAGE) and PRPF39 as prey (top, western blot with anti-PRPF39 antibody) shows weak binding when compared to the GST control.