Research Paper Volume 12, Issue 10 pp 8893—8912

Long noncoding RNA lnc-LOC645166 promotes adriamycin resistance via NF-κB/GATA3 axis in breast cancer

Figure 6. lnc-LOC645166 recruits NF-κB to promote GATA3 transcription. (A) RNA pulldown assays were performed and the interaction between lnc-LOC645166 and NF-κB was confirmed by Western blot. (B) RIP assay of the enrichment of NF-κB with lnc-LOC645166 relative to IgG in the lysates of 231/ADR and MCF-7/ADR cells (n =3). (C) NF-κB motif. (D) The correlation between lnc-LOC645166 and GATA3 expression was assessed in breast cancer tissues using a Pearson’s correlation analysis. (E) The correlation between NF-κB and GATA3 expression was assessed in TCGA Breast Cancer samples using a Pearson’s correlation analysis. (F) Representative immunostaining of NF-κB and GATA3 in tumor samples with good or poor responses to adriamycin therapy. R means resistant to ADR therapy, S means sensitive to ADR therapy. (G) RT–qPCR analysis of GATA3 in breast cancer cells transfected with pcDNA-LOC645166 or cotransfected with si-NF-κB. (H) RT–qPCR analysis of GATA3 in adriamycin resistant cells transfected with sh-LOC645166 or cotransfected with pcDNA-NF-κB. (I) Chromatin immunoprecipitation (ChIP) assays were performed to determine the affinity of NF-κB on the promoter region of the GATA3 locus after lnc-LOC645166 overexpression or knockdown. 2% Input cell lysate was used data are shown as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.