Fig. 6: BRG1 mediates the epigenetic loss of HLF.

a Schematic showing the generation of primary tumor cells (786-O-P) and lung metastatic cells (786-O-LM). Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. b–d RT-qPCR (n = 3 biological replicates) (b), immunoblotting (c) (repeated at least two times independently with similar results), representative images and quantification of 3D spheroid invasion (d), n = 6 independent cell cultures. Scale bar, 200 µm. e Representative images and quantification of bioluminescence imaging post tail vein inoculation into NSG mice (n = 8 mice for 786-O-P, n = 10 mice for 786-O-LM). f ChIP-seq data of H3K27ac and H3K4me1 and ATAC-seq data at chr17:53,334,969-53,405,254 (hg19). The targeting sites of sgHLFs (used in i) at the enhancer region are indicated. g Immunoblotting in cells treated with C70 (20 µM) or vorinostat (2 µM) for 24 h. This experiment has been repeated three times with similar results. The samples derive from the same experiment but different gels for each target protein were processed in parallel. h Schematic showing CRISPR interference system (dCas9-KRAB) and CRISPR activation system (dCas9-p300). Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. i RT-qPCR in cells applied with dCas9-KRAB system (left) and dCas9-p300 system (right) respectively targeting specific HLF enhancer region, n = 3 biological replicates. j Potential regulators binding at the validated regulatory enhancer region of HLF. k Immunoblotting in cells transduced with shCtrl or SMARCA4 shRNAs. This experiment has been repeated three times with similar results. The samples derive from the same experiment but different gels for BRG1, Vinculin and another for FLAG were processed in parallel. l Schematic showing the functional mechanism of AU-15330 that can degrade BRG1. Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. m, n Immunoblotting (m) and RT-qPCR (n = 3 biological replicates) (n) in cells treated with AU-15330 (0.1 µM, 1 µM) for 48 h. Representative immunoblots shown in m were repeated at least two times independently with similar results. For A375 MA2 and HCT116 cells, the samples derive from the same experiment but different gels for BRG1 and Vinculin were processed in parallel. o, p Representative images (o) of primary tumors (PT) and lung metastatic tumors (Lung-met), and signals of Lung-Met relative to its matching PT (p) from orthotopic injection in NSG mice, n = 6 mice in Ctrl and shSMARCA4 group, n = 5 mice in rescue group. Data are mean ± s.e.m. (b, e, i, n), box plots show the median and interquartile range, and whiskers show the data range (d, p). One-way ANOVA followed by a post hoc Dunnett-t-test (i, n, p) or unpaired two-tailed t-test (b, d, e). Source data are provided as a Source Data file.