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. 2022 Sep 20;119(39):e2117988119. doi: 10.1073/pnas.2117988119

Fig. 4.

Fig. 4.

TGFB2-AS1 interacts with SMARCA4. (A) RNA-pulldown assay to identify TGFB2-AS1 binding proteins in MDA-231 cells. The arrow and red bracket indicate the band representing the TGFB2-AS1 specific binding protein identified by mass spectrometry as SMARCA4. (B) Western blot identifying SMARCA4, BAF170, and SNF5 present in TGFB2-AS1 pull-down precipitates with Lamin B1 and β-tubulin as negative controls. (C–E) The interaction of TGFB2-AS1 with SMARCA4 (C, a semiquantitative RT-PCR was inserted), BAF170 (D), and SNF5 (E) was verified by an RIP assay together with U6 and GAPDH as negative controls. (F) Schematic representation of Flag-tagged full-length human SMARCA4 and its deletion mutants. (G) The anti-Flag Western blot images showing expressions of full-length or deleted SMARCA4 in MDA-231 cells. (H) RIP and subsequent RT-PCR assays in MDA-231 for TGFB2-AS1 enrichment of human full-length SMARCA4 and its deletion mutants. (I) Schematic representation of full-length and various truncated TGFB2-AS1. (J) Western blot showing SMARCA4, BAF170, and SNF5 pulled down by full-length and various truncated TGFB2-AS1. Error bars represent mean ± SD (n = 3). Statistical significance was assessed using two-tailed Student’s t test. ***P < 0.001.