Fig. 4: HDAC2 interacts with the TGF-β related proteins, SMAD3-SKI in BTSCs.

a Kaplan–Meier survival curves for mice orthotopically xenografted with single and double HDAC1/2 KO BT67 cells compared to AAVS1 control mice (Log-rank (Mantel–Cox) method, **p < 0.01, ****p < 0.0001, n = 10). b Kaplan–Meier survival curves for mice orthotopically xenografted with HDAC2 KD BT147 cells compared to mice xenografted with their respective scrambled control cells (Log-rank (Mantel–Cox) method, ****p < 0.0002, n = 10). c–f Co-immunoprecipitation and sequential co-IP assays in dual crosslinked BT67 cells. (n = 3). g In situ proximity ligation assays (PLA) confirm protein-protein interactions between HDAC2 and the SMAD3-SKI complex which was disrupted following 24 h treatment with romidepsin relative to DMSO control in BTSCs. Quantitative representation of the average number of PLA foci counted in 30 nuclei/reaction in BT67. Significance was determined using ANOVA (Tukey’s test) at 95% confidence intervals, **p < 0.01, ***p < 0.001; Data are represented as mean values ± SEM; n = 3. HDAC1/2 protein-protein interaction was used as positive control and PLA probe only and IgG only were used as negative controls for the assay. h, i Co-IP assays in dual crosslinked HEK293T/17 cells expressing different SMAD3 (SMAD3, SMAD3NL, SMAD3C and SMAD3ΔC) and HDAC2(HDAC2N and HDAC2C) mutants or empty vector (n = 3 biologically independent transductions). Input: 1% input was used for all the co-IP assays. IP-immunoprecipitation, ID-immunodepleted samples, IgG- antibody controls for non-specific pull down. - represents molecular weight markers (50 and 150). Source data are provided in the source data file.