Fig 1: Reprogramming of HSCs by DNMT3B suppression. (A) Expression levels of DNMT family and Smad2/3-related proteins in Huh7 (left) and LX2 cells (right) were examined after treatment with 20 ng/ml TGF-β1. (B) Size and morphology of MCHSs composed of Huh7 and DNMT3B-deficient LX2 cells (in a 7:3 ratio) for 3 days. Two different siRNA used (upper: purchased from Dharmacon, lower: purchased from Bioneer). (C) DNMT3B-deficient LX2 cells were treated with or without 20 ng/ml TGF-β1 for 48 h. The morphology and α-SMA intensity were examined and calculated using an Operetta HTS system and Harmony software. (D) EMT-related proteins were examined in DNMT3B-deficient LX2 cells with or without 20 ng/ml TGF-β1 treatment. (E) Size and morphology of MCHSs composed of Huh7 and LX2 (in a 7:3 ratio) treated with 10 uM nanaomycin A for 3 days. (F) EMT-related proteins were examined in nanaomycin A treated LX2 cells with or without 20 ng/ml TGF- β1 treatment. β-actin was used as a control.
Fig 2: mRNA relative expression levels of altered genes involved in the DNA methylation process in ischaemic hearts versus controls. (a) Methylation machinery of DNA: genes related to addition (DNMT3B), elimination (TET1 and SMUG1) and maintenance (MBD2 and UHRF1) of methyl groups to DNA. (b) Methylation machinery regulators (AHCY, TRAF6, GADD45G and OGT). The results were obtained by mRNA-sequencing SOLiD 5500XL platform. Data are presented as the mean ± SEM. a.u., arbitrary units. Ischaemic cardiomyopathy patients (n = 13; blue), controls subjects (n = 10; grey). * p < 0.05, ** p < 0.01, *** p < 0.001.
Fig 3: The efficacy of chemotherapeutic drugs against HCC was increased by CHIR-99021. (A) MCHS size was examined after treatment with 1 or 2 µM sorafenib with or without 0.5 µM CHIR-99021 for 2 days. Images (upper) were obtained using an Operetta HCS system, and MCHS sizes were calculated using Harmony software. (B) Cleaved-caspase 3, an apoptosis marker, was examined in MCHSs after treatment with 1 or 3 µM sorafenib with or without 0.5 µM CHIR-99021. (C) Tumor volume was measured for 3 weeks (1 week before and 2 weeks after drug administration) in the 10 mpk sorafenib, 15 mpk CHIR-99021, and 10 mpk sorafenib plus 15 mpk CHIR-99021 groups. (D) Liver toxicity was determined by examining mouse blood (AST; upper, ALT; lower) after completing the in vivo experiment. (E) Expression levels of DNMT3B and Cleaved PARP, an apoptosis marker, were calculated in xenografted tumor tissue. β-actin was used as a control.
Supplier Page from Abcam for DNMT3B Assay Kit