7 research outputs found
<i>Arid4b</i> germline amino acid sequence variants.
<p>Substitutions in the AKR allele are shown relative to a consensus sequence that is identical between DBA, FVB, and C57Bl/6. The+symbol indicates conserved substitutions.</p
Cellular processes regulated by <i>Arid4b</i> knockdown.
<p>Biological processes are listed in order of statistical significance after applying a cutoff based on z-scores with an absolute value greater than 2. Negative z-scores indicate downregulation; positive z-scores indicate upregulation.</p
High expression of <i>ARID4B</i> is associated with poor clinical outcome.
<p>In patients with ER-positive tumors who were node negative at diagnosis (A) distant metastasis-free survival (DMFS) was significantly lower (pâ=â.009) in patients with high expression (blue) compared to middle (red) or low (gray) expression of <i>ARID4B</i>, and multivariate analysis of 440 cases (B) was performed to determine metastatic progression hazard ratios of 0.54 and 0.42 for median and low <i>ARID4B</i> terciles, respectively, compared to the high <i>ARID4B</i> tercile. The association of high <i>ARID4B</i> with poor DMFS was also highly significant (pâ=â3.05Ă10â§â4) among ER-positive patients in the absence of adjuvant therapy (C) with similar hazard ratios (D) of 0.53 and 0.49 for middle and low <i>ARID4B</i> groups compared to high <i>ARID4B</i>.</p
<i>Arid4b</i> expression is correlated with tumor growth and metastasis.
<p>Gene expression microarray data from 18 of the AKXD recombinant inbred strains shows expression of <i>Arid4b</i> is positively correlated with tumor burden (A) and metastatic density (B). The formula used to calculate metastatic density was ln[(metastases/um<sup>2</sup>)Ă10<sup>8</sup>].</p
ARID4B binds BRMS1.
<p>HA-BRMS1 was coprecipitated with FLAG-ARID4B (A) and FLAG-ARID4B was coprecipitated with myc-BRMS1 (B). No difference in binding to BRMS1 was observed between the AKR and DBA alleles of ARID4B.</p
Stable shRNAâmediated knockdown of <i>Arid4b</i> inhibits lung metastasis.
<p>ARID4B protein levels were analyzed by western blot to determine level of knockdown in cell lines stably expressing different <i>Arid4b</i> shRNAs. Experiment was performed in triplicate and a representative blot is shown in panel A. Lysate from 293 cells transiently transfected with <i>Arid4b</i> was used as a positive control (+C). Untransduced 6DT1 and scrambled control lines are designated âuntrâ and âscrâ respectively. The lower non-specific (n.s.) band most likely represents ARID4A, based on a nearly identical epitope sequence and consistent with the observed difference in molecular weight. Expression of the non-specific band remained relatively unchanged across the panel of stable lines. Densitometry was performed to quantitate knockdown at the protein level in the <i>Arid4b</i> shRNA lines relative to the scrambled control line (B) Data are shown as the mean ± standard error of three independent experiments. Kruskal-Wallis followed by Conover-Inman post hoc tests were used to determine statistical significance versus the scrambled control cell line. *, p<.05; **, p<.01; ***, p<.001. Primary tumors were weighed (C) and lung surface metastases were counted (D) at 3.5 weeks after orthotopic implantation of 10â§5 cells from the scrambled control, H3, and H4 lines into the mammary fat pad of FVB mice (nâ=â10 per cohort). Horizontal bars represent median values and statistical significance was determined using Kruskal-Wallis tests followed by Conover-Inman correction for multiple comparisons.</p
Differential binding of the AKR and DBA variants of ARID4B to the mSIN3A complex.
<p>Western blots (A) show equivalent input of V5-ARID4B, mSIN3A, mSDS3, and equivalent pull-down of the two variants using an anti-V5 antibody. Decreased binding of the DBA variant of ARID4B to mSIN3A and mSDS3 was observed relative to the AKR variant. Experiment was performed in triplicate and densitometry analysis (B) revealed a 51% reduction in mSIN3A binding (pâ=â.037) and a 37% reduction in mSDS3 binding (pâ=â.026) for the DBA variant relative to AKR. Statistical significance was determined using paired t-tests. *, p<.05.</p