Journal: bioRxiv

Article Title: Analysis of Gene Expression Changes upon Topobexin Treatment and TOP2B-knockout in hiPSC derived cardiomyocytes

doi: 10.64898/2026.03.09.710520

Figure Lengend Snippet: (A) The differentiation protocol resulted in the conversion of monolayers of wild type (WT) or TOP2B null (line 18D, BKO) hiPSC cells into layers of beating cardiomyocytes in 8-12 days. However, on average this took 1 day longer for BKO than WT hiPSC cells. Each spot represents a separate hiPSC to CM differentiation experiment, four biological replicates for WT and five biological replicates for the BKO. Line = mean. Statistical significance was determined using a two-tailed unpaired t-test, * - P < 0.05. (B) Primary component analysis of RNA-seq analysis comparing WT and TOP2B null (BKO) hiPSC and derived CM (quadruplicate biological replicas). (C) Volcano plots to visualise gene expression changes upon differentiation of WT and BKO hiPSC cells to CM. Red lines represent cutoffs of > 2-fold change in transcript abundance with a P adj of <0.05. Numbers in the corners of the plots highlight the number of up or down regulated protein-coding genes in the CM compared to the hiPSC cells, using the above cutoffs.

Article Snippet: The next day, they were incubated with primary anti-TOP2B antibody (MAB6348, 1:500, R&D Systems) and anti-β-actin antibody (NB600-501, 1:5000, Novusbio) for 1 h at room temperature, then washed with 0.1% TBS-T (2× 1 min, 1× 10 min, repeated twice).

Techniques: Two Tailed Test, RNA Sequencing, Derivative Assay, Gene Expression

Journal: bioRxiv

Article Title: Analysis of Gene Expression Changes upon Topobexin Treatment and TOP2B-knockout in hiPSC derived cardiomyocytes

doi: 10.64898/2026.03.09.710520

Figure Lengend Snippet: (A) Volcano plot illustrating the number of genes up- or down regulated in BKO versus WT CM. (B) Effect of topobexin gene expression (transcript abundance) in WT CM. (C) Effect of ICRF-187 on gene expression (transcript abundance) in WT CM. For volcano plots, cut-offs (red-lines) were as in . Representative genes are highlighted. (D) Primary component analysis of transcript abundance comparing in four conditions, WT and TOP2B null (BKO) CM, WT CM treated with topobexin and CM treated with ICRF-187 (quadruplicate biological replicas).

Article Snippet: The next day, they were incubated with primary anti-TOP2B antibody (MAB6348, 1:500, R&D Systems) and anti-β-actin antibody (NB600-501, 1:5000, Novusbio) for 1 h at room temperature, then washed with 0.1% TBS-T (2× 1 min, 1× 10 min, repeated twice).

Techniques: Gene Expression

Journal: bioRxiv

Article Title: Analysis of Gene Expression Changes upon Topobexin Treatment and TOP2B-knockout in hiPSC derived cardiomyocytes

doi: 10.64898/2026.03.09.710520

Figure Lengend Snippet: Differential gene expression tables were filtered to include only genes that change significantly ( P adj <0.05) in expression in both TOP2B null CM vs WT cells and after treatment of WT CM with topobexin. The gene list was further filtered to only include protein-coding genes. Log2 FC values are plotted on the x and y axes. r = Pearson corelation coefficient.

Article Snippet: The next day, they were incubated with primary anti-TOP2B antibody (MAB6348, 1:500, R&D Systems) and anti-β-actin antibody (NB600-501, 1:5000, Novusbio) for 1 h at room temperature, then washed with 0.1% TBS-T (2× 1 min, 1× 10 min, repeated twice).

Techniques: Gene Expression, Expressing

Journal: bioRxiv

Article Title: Analysis of Gene Expression Changes upon Topobexin Treatment and TOP2B-knockout in hiPSC derived cardiomyocytes

doi: 10.64898/2026.03.09.710520

Figure Lengend Snippet:

Article Snippet: The next day, they were incubated with primary anti-TOP2B antibody (MAB6348, 1:500, R&D Systems) and anti-β-actin antibody (NB600-501, 1:5000, Novusbio) for 1 h at room temperature, then washed with 0.1% TBS-T (2× 1 min, 1× 10 min, repeated twice).

Techniques: