Fig. 3: The MLL core complex is required for transcriptional activation by FOXQ1.
From: FOXQ1 recruits the MLL complex to activate transcription of EMT and promote breast cancer metastasis
a Validation of a panel of FOXQ1 upregulated genes in HMLE/FOXQ1cells. CTSB and IL17RA serve as RbBP5-independent controls. Data were analyzed by 2−ΔΔCT method with internal sample normalization against β-actin and compared to the HMLE/LACZ control cells (n = 3). b The effects of RbBP5 knockdown (KD) on the expression of a panel of FOXQ1-RbBP5 EMT targets in HMLE/FOXQ1 cells. Results are relative to HMLE/FOXQ1 nontarget (NT) control cells normalized to β-actin. c The effects of KMT2A/MLL1 silencing on the expression of EMT target genes were assessed by qRT-PCR of HMLE/FOXQ1 shMLL1 cells. Results are relative to HMLE/FOXQ1 nontarget (NT) control cells normalized to β-actin. d The effects of OICR-9429 treatment on the expression of the FOXQ1-RbBP5 EMT gene panel were evaluated by qRT-PCR in HMLE/FOXQ1 cells. Results are relative to DMSO mock treatment and normalized to β-actin. e The impact of RbBP5 KD on FOXQ1 target promoter occupancy was assessed using V5-FOXQ1 ChIP-qPCR from HMLE/FOXQ1 cells with shRbBP5 or NT control. Signals were normalized to the input chromatin sample. f The impact of RbBP5 KD on H3K4me3 levels within the promoters of FOXQ1 target genes was evaluated via H3K4me3 ChIP-qPCR from HMLE/FOXQ1 cells with shRbBP5 or NT control. Results were evaluated by qPCR, and the data were normalized to a matched DNA sample from 1% input chromatin. g The impact of FOXQ1 on RbBP5 occupancy within EMT promoter regions was evaluated via RbBP5 ChIP-qPCR in HMLE/FOXQ1 cells with shFOXQ1 or NT control. Results were analyzed using the same approach as panel f. h The effects of FOXQ1 depletion on H3K4me3 levels within EMT promoter regions were assessed by H3K4me3 ChIP-qPCR in HMLE/FOXQ1 cells with FOXQ1 shRNA or NT control. Results were analyzed using the same approach as panel f. For a–h panels, bars indicate the mean ± SD (n = 3). Dots indicate the individual replicate values normalized to the control group’s mean. Results were analyzed by unpaired, two-tailed t-test with Bonferroni multiple comparison adjustment. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided in Source data 1.
