Fig. 8: Increased pY54-H3 marks is associated with NSML.

a Lysates from prostate, testis, and liver of NSML/+ mice were immunoprecipitated with pY54-H3 antibodies, followed by immunoblotting with H3 antibodies (top panel). Lysates were also subjected to immunoblotting with actin antibodies (lower panel). b ChIP-seq was performed using pY54-H3 antibody and the peaks at enhancer (AR1) and exon 1 (AR2) sites of the mouse Ar are shown in graphical format. The numbers indicate nucleotide position at the midpoint of the peaks. c Lysates from prostate, testis and liver of NSML/+ (LS) mice were subjected to ChIP using pY54-H3 (or IgG as control) antibody, followed by qPCR using primers corresponding to AR regions 1 and 2. (n = 3 mice in each group, 3 replicates). d, e Total RNA isolated from prostate, testis and livers of NSML/+ (LS) mice and were subjected to qRT-PCR with AR, TMPRSS2 and actin primers. (n = 4 mice in each group, 3 replicates). f iPSCs derived from a NSML patient (Q510E iPSC) and the healthy individual (WT iPSCs) were subjected to IP with pY54-H3 antibody, followed by immunoblotting with H3 antibodies (top panel). Lower panels are immunoblots with the indicated antibodies. g iPSCs (derived from a NSML patient) lysates were subjected to ChIP using pY54-H3 antibody (or IgG), followed by qPCR using primers corresponding to AR exon 1 and 2. For a and f, representative images are shown from 3 biologically independent experiments. For c, d, e, g, data are represented as mean ± SEM. n = 3 biologically independent samples (three replicates). 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. h Graphical Abstract: SHP2 phosphorylation by ACK1 promotes interaction with AR, followed by nuclear translocation. SHP2 erases pY54-H3 epigenetic marks upregulating AR transcription, a schematic representation.