Journal: Stem cell research & therapy
Article Title: Functional analysis of JPH2-knockout cardiomyocytes identifies ECCD as a novel indicator in a human cardiac modelJPH2.
doi: 10.1186/s13287-025-04323-4
Figure Lengend Snippet: Fig. 1 Generation and characterization of JPH2-knockout hESC-derived cardiomyocytes. (A) Schematic of the sgRNA targeting the MORN region 2 of JPH2, resulting in a 16-bp deletion from 333 amino acids. (B) Flow cytometry analysis of cardiomyocyte differentiation efficiency at day 15 post- differentiation. (C) Bar graph showing the percentage of TNNT2-positive cells in WT-CMs and KO-CMs. (D) Morphology of WT-CMs and KO-CMs at day 15 post-differentiation (40x magnification). (E) Western blot analysis confirming the knockout of JPH2 protein in KO-CMs. Full-length blots are presented in Figure S8. (F) Immunofluorescence staining of ventricular-specific marker MYL2 and atrial-specific marker MYL7 in WT-CMs and KO-CMs: MYL2 in green, MYL7 in red, DAPI in blue. Scale bar = 100 μm. Results are presented as means ± SEMs of 3 independent experiments. ns, not significant, unpaired 2-sided Student’s t test
Article Snippet: For atrial and ventricular specific markers identification, cardiomyocytes were incubated overnight at 4 °C with 1:200 rabbit monoclonal IgG MYL7 (Abcam, ab205374) and 1:200 mouse IgG1κ MYL2 (Santa Cruz, sc-517244).
Techniques: Knock-Out, Derivative Assay, Flow Cytometry, Western Blot, Immunofluorescence, Staining, Marker