Fig 2: dCas9-SALL1-SDS3 recruits the SIN3A complex. (A, B) Volcano plots of enriched protein interactions in dCas9-SALL1-SDS3 versus WT Co-IPs (A) or versus dCas9 Co-IPs (B), identified by mass spectrometry. Hits were defined as interactions enriched >2.5-fold (log2-fold-change >1.32) in dCas9-SALL1-SDS3 Co-IPs at a significance threshold of p < 0.0008 (adjusted p-value <0.05), denoted by vertical and horizontal dashed lines, respectively. n = 4 (dCas9-SALL1-SDS3 and dCas9) or n = 3 (WT) biological replicates per group. dCas9 and dCas9-SALL1-SDS3 are denoted in red. (C) Overlap of significantly enriched protein interactions between dCas9-SALL1-SDS3 versus WT and dCas9-SALL1-SDS3 versus dCas9 Co-IP comparisons. White text denotes components of the Sin3 complex. (D) Co-IP/Western blot analysis using nuclear extracts from U2OS cells stably expressing FLAG-tagged dCas9, dCas9-SALL1-SDS3, or dCas9-KRAB. Following Co-IP with α-FLAG, proteins were resolved by SDS-PAGE and immunoblotted for HDAC1, HDAC2, RBBP4, and FLAG. n = 5 biological replicates per group. Co-IP, Co-immunoprecipitation; SDS-PAGE, sodium dodecyl-sulfate polyacrylamide gel electrophoresis; WT, wild-type.

Fig 3: ARMC12 interacts with RBBP4 protein in NB cells. a Co-IP, Commassie blue staining (left panel) and mass spectrometry (MS) assay (right panel) showing the identification of ARMC12-binding protein in SH-SY5Y cells stably transfected with empty vector (mock) or ARMC12. b Co-IP and western blot assays indicating the endogenous interaction between ARMC12 and RBBP4 protein in BE(2)-C and IMR32 cells. The immunoglobulin G (IgG)-bound protein was taken as negative control. c Co-IP and western blot assays (lower panel) depicting the interaction between ARMC12 and RBBP4 protein in SH-SY5Y cells transfected with mock, FLAG-tagged ARMC12 truncates (upper panel), and HA-tagged RBBP4. d Co-IP and western blot assays (lower panel) showing the interaction between ARMC12 and RBBP4 protein in SH-SY5Y cells transfected with mock, HA-tagged RBBP4 truncates (upper panel), and FLAG-tagged ARMC12. e, f Co-IP and western blot (e, middle and right panels), and BiFC (f, arrowheads) assays indicating the interaction between ARMC12 and RBBP4 protein in SH-SY5Y cells transfected with wild-type or mutant (e, left panel) FLAG-tagged ARMC12 and HA-tagged RBBP4. Scale bars: 10 μm. Data are representative of three independent experiments

Fig 4: Analysis of the mechanism regulating microglial polarization by GAS5 ASilver staining to identify specific binding partners of GAS5 after RNA pull‐down experiment.BRNA IP analysis between GR and GAS5. N = 3 experiments.CRNA IP analysis between RbAp48 and GAS5. N = 3 experiments.DRNA IP analysis between RING1A/B and GAS5. N = 3 experiments.E–GChIP analysis of microglia transduced with the CtrlOE or GAS5OE lentivirus. The promoter regions of STAT6 (E), PPARγ (F) and CEBP/β (G) were detected in MGGAS5OE versus the control using the anti‐EZH2 antibody.Data information: ***P < 0.001 (B–D, Student's t‐test). Data are shown as the mean ± SD.

Fig 5: RbAp48 in combination with radiation inhibits cell proliferation and results in G2 cell cycle arrest. (A) An MTT assay was performed to assess AGS cell proliferation. (B) The percentage of cells in each phase was determined by (C) flow cytometry analysis. *P<0.05, **P<0.01, ***P<0.001 vs. control; ^P<0.05, ^^P<0.01, vs. mock; &P<0.05 vs. control+RAD; #P<0.05 vs. mock+RAD. RbAp48, retinoblastoma-binding protein 4; control, PBS; mock, pcDNA3.1; RAD, 6 Gy radiation.