Fig. 3: SHP2 erases pY54-H3 epigenetic marks at the AR gene locus.

a–c VCaP, C4-2B and LAPC-4 cells were treated with 1 μM of (R)−9b or SHP099 for 8 h and lysates were subjected to ChIP with pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon 1, 2, or control (IGX, gene desert on chromosome 12) region. d VCaP cells were co-transfected with HA-tagged ACK1 or kdACK1 expressing constructs and lysates were subjected to ChIP using pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon 1, 2 or control region. e VCaP cells were co-transfected with HA-tagged ACK1 and FLAG-tagged SHP2 or SHP2-Y62F, -Y279F or -Y580F mutants. The lysates were subjected to ChIP using pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon 1, 2 or control region. f LAPC-4 cells were transfected with FLAG-tagged H3 or H3-Y54F mutant and treated with 1 μM of (R)−9b or SHP099 for 8 h and lysates were subjected to ChIP with pY54-H3 antibodies, followed by qPCR using primers corresponding to AR exon 1, 2, or control region. g LAPC4 cells were treated with 1 μM of Enz or Abr for 8 h and ChIP was performed using pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon1, 2 or control region. h LAPC4 cells were co-transfected with HA-tagged ACK1 and FLAG-tagged AR or AR-Y267F, -Y307F, -Y534F, -Y223/363 F expressing constructs. ChIP was performed using pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon 1, 2 or control region. i C4-2B cells were transfected TNK2, AR and PTPN11 siRNA and ChIP was performed using pY54-H3 antibody, followed by qPCR using primers corresponding to AR exon 1, 2 or control region. For a–i, data are represented as mean ± SEM (n = 3 biologically independent experiments, three replicates). For a–c, e–i, p values were determined by one-way ANOVA. For d, p values were determined by unpaired two-tailed Student’s t-test. p values are shown on the graph. Source data are provided as a Source Data file.