Normal rabbit igg

qChIP assays were performed by incubating chromatin with antibodies against Ikaros15 (link)16 (link)17 (link), H3K9me3 (Abcam, Cambridge, MA, USA) or normal rabbit IgG (Abcam) as a control15 (link)16 (link)17 (link)48 (link). Enrichment of the ChIP sample over input was evaluated by qPCR with ≥ 3 replicates using specific primers in the promoter region of DNM2(forward: 5′-ACCGCGGGATGGAAGAG-3′, reverse: 5′-TGAAGGCGTCCTGCAGTTT-3′). Relative concentration of the qPCR product is presented as the fold change of the level of DNA-Ikaros and DNA-H3K9me3 samples compared with controls.

The hUC‐MSCs were culture in osteogenic‐induced medium, after 7 days cross‐linked with 1% formaldehyde at 37°C for 10 minutes, terminated with 0.125 mol/L Glycine (Bioshrap) for 10 minutes. Then, 2 × 10⁷cells were harvested and lysed in SDS lysis buffer supplemented with cocktail. The cell lysis was sonicated with Cole‐Parmer Instruments CP130 (Vernon) to shear chromatin, then centrifuged at 13 000 g for 15 minutes. The supernatants fixed with antiSTAT3‐antibody (Cell Signaling Technology), c‐JUN (Cell Signaling Technology), and normal rabbit IgG (Abcam), adding Protein‐A/G Immunoprecipitation Magnetic Beads (Biomake) for overnight at 4.0°C. After interacting 10% Chelex‐100 mixtures (Bio‐Rad) to reverse crosslinking, DNA was purified and examined by qPCR. The ChIP‐qPCR primer sequences are included in Table S1, and all samples were performed in triplicate independent tests.

According to the manufacturer’s instructions, ChIP assays were performed using the ChIP Assay Kit (Beyotime Biotechnology). Briefly, the chromatin complexes were treated with anti-PRRX2 antibody or normal rabbit IgG (Abcam). Then the immunoprecipitated DNA was extracted and purified by qPCR. The primers for ChIP qPCR are listed in Table S3.

The qChIP assays were performed by incubation of the chromatin with antibodies against IKZF1 [14 (link)–16 (link)], H3K9me3 (Abcam, USA) or normal rabbit IgG (Abcam) as a control [14 (link)–16 (link), 23 (link)]. Enrichment of the ChIP sample over input was evaluated by qPCR with three or more replicates, using specific primers in the promoter region of CRLF2 as shown in Supplementary Table 3 (forward: 5′-AGGGGA AGGGAGGGGAAGGGAAAG-3′, reverse: 5′-CCCTTTCCTCCCCTCCCCTCCT-3′). Relative concen- tration of the qPCR product was presented as the fold change of the level of DNA-IKZF1 and DNA-H3K9me3 samples in comparison to controls.