Figure 3. LINC00839 serves as a scaffold to recruit Ruvb1 to the Tip60 complex and increases its acetylase activity.

• A
  An RNA pull‐down assay was performed to identify proteins that bind to LINC00839 in HCT116 cells.
• B
  RNA pull‐down assay and WB showed that biotinylated LINC00839 can bind to Ruvb1 in HCT116 cells. Dot blot of RNA–protein interactions indicating equal amounts of RNA used in the assay.
• C
  RIP experiments were performed to confirm the binding of LINC00839 to Ruvb1. IgG was used as a negative control antibody. Primers specific for U6 and GAPDH were used as negative control primers.
• D
  Different fragments of LINC00839 (solid line) and its antisense sequence (dotted line) were used for the RNA pull‐down assay and the subsequent WB.
• E
  Full‐length LINC00839, antisense sequence of LINC00839, and LINC00839 sequence lacking nucleotides 1,033–1,290 were used for the RNA pull‐down assay and the subsequent WB.
• F
  RIP experiments were performed by incubating Tip60, INO80, SRCAP, and PIH1D1 antibodies with HCT116 cell lysates.
• G
  The effect of LINC00839 on the expression of Tip60 and Ruvb1 was determined by WB.
• H
  Interaction of endogenous Ruvb1 and Tip60 in CRC cells was confirmed by Co‐IP.
• I
  The interaction between Ruvb1 and Tip60 in cells overexpressing wild‐type LINC00839 and LINC‐Δ5 was confirmed by Co‐IP.
• J
  The interaction between Ruvb1 and Tip60 in LINC00839‐KD cells and cells expressing LINC‐Δ5 was confirmed by Co‐IP.
• K
  Acetylase activity assays were performed with LINC00839‐overexpressing and LINC00839‐knockdown cells.
• L
  Acetylase activity was measured in SW480 and LoVo cells subjected to different treatments.

Data information: In (C, F, K and L) the data are presented as the mean ± SD and were analyzed by unpaired t‐test, n = 3 technical replicates. Across experiments, the P‐values are as follows: ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Source data are available online for this figure.