Journal: bioRxiv

Article Title: Increased utrophin expression in healthy and DMD patient derived myoblasts in response to ERK1/2 and EZH2 inhibitor treatment

doi: 10.64898/2026.04.13.718206

Figure Lengend Snippet: Corresponding to . a . Basal levels of MYOD1 mRNA in primary healthy human myoblasts (n=6) and primary DMD patient-derived myoblasts (n=16). Lines show median and quartiles. b . AB1098 cells were treated with 10 μM LY32 +/- 10 μM GSK503 or vehicle (DMSO) for 24 hours. Directly following treatment (0 days since drug removal) cells were manually counted, with trypan blue staining to discount dead cells. Cells were then replated at a constant concentration, cultured in proliferation media without drugs for 24 hours and counted again (1 day since drug removal). Cells were then replated at a constant concentration and cultured again for 6 days before counting (7 days since drug removal). Cells were passaged once in this 6-day period. Cell count is shown relative to the vehicle only control for each time point. c . RT-qPCR of MYOD1 in immortalised myoblasts AB1190 and AB1098 following treatment with varying concentrations of Ruxolitinib (Ruxo). Results are shown relative to vehicle treated sample with a two-way ANOVA with Dunnetts multiple comparison test to determine significance. Graph shows mean (n=3) +/- SEM. a-b . MYOD1 is normalised against TBP . *p<0.05, ***p<0.001, ****p<0.0001.

Article Snippet: The muscle biopsy was enzymatically digested in basal medium (C-23060; PromoCell, Heidelberg, Germany) containing collagenase D (2 mg/mL, Roche, Germany) and Dispase II (2 mg/mL, Sigma) for 1 h at 37°C with trituration every 15 min. After filtering through a 100-μm filter (BD Falcon) and centrifugation cells were resuspended in proliferation medium (15% FBS, C-23060; PromoCell, Heidelberg, Germany) and transferred to a T-25 tissue culture vessel (Nunc, Germany).

Techniques: Derivative Assay, Staining, Concentration Assay, Cell Culture, Cell Characterization, Control, Quantitative RT-PCR, Comparison