ventricular cardiomyocyte kit  (Axol Bioscience)

Journal: bioRxiv

Article Title: Cardiac defects in spinal muscular atrophy and the role of SMN in cardiomyocyte homeostasis

doi: 10.64898/2026.03.20.713246

Figure Lengend Snippet: KO schematic in cardiomyocytes B-D: Oxygen consumption rate (OCR) is significantly increased in siSMN-treated cardiomyocytes compared with siScramble controls and extracellular acidification rate (ECAR) is significantly decreased in siSMN-treated cardiomyocytes. n = 16 technical replicates. * p <0.05. **p < 0.01. Mitochondrial and glycolytic ATP production rates show no significant difference between siSMN and siScramble conditions (ns). E: Volcano plot of differential gene expression following SMN knockdown. F: Heatmap and hierarchical clustering of differentially expressed genes demonstrate distinct transcriptional profiles between siSMN and siScramble cardiomyocytes. G: Pathway enrichment analysis of differentially expressed genes identifies significant perturbation of multiple signaling pathways, including enrichment of PTEN signaling.

Article Snippet: Human cardiomyocytes (Axol Bioscience Limited, Cambridgeshire, England; Cat. No. ax2520; Lot No. 2520310317) were cultured and differentiated in coated plates using supplemented media from the Human iPSC-Derived Ventricular Cardiomyocyte Kit (Axol Bioscience Limited, Cambridgeshire, England).

Techniques: Gene Expression, Knockdown, Protein-Protein interactions

Journal: bioRxiv

Article Title: Cardiac defects in spinal muscular atrophy and the role of SMN in cardiomyocyte homeostasis

doi: 10.64898/2026.03.20.713246

Figure Lengend Snippet: Ingenuity pathway analysis of differentially expressed genes after SMN2 knockdown in human cardiomyocytes, showing the top 20 significantly enriched canonical pathways ranked by –log(p value). Bar color denotes predicted directionality based on z-score: black indicates negative z-score (predicted pathway inhibition), red indicates positive z-score (predicted pathway activation), and gray indicates no consistent activation pattern.

Article Snippet: Human cardiomyocytes (Axol Bioscience Limited, Cambridgeshire, England; Cat. No. ax2520; Lot No. 2520310317) were cultured and differentiated in coated plates using supplemented media from the Human iPSC-Derived Ventricular Cardiomyocyte Kit (Axol Bioscience Limited, Cambridgeshire, England).

Techniques: Knockdown, Inhibition, Activation Assay

Journal: iScience

Article Title: Crosstalk between circulating DPP3 and immune cells in the context of cardio-systemic stress

doi: 10.1016/j.isci.2026.115114

Figure Lengend Snippet: A fraction of DPP3 is transported via EVs and upregulated during septic shock in humans (A) Western blot qualitative analysis of cluster of differentiation 63 (CD63) at 26 KD (bottom), heat shock protein 70 (HSC70) at 73 KD (middle), and DPP3 at 83 KD (above), in protein extracts of EVs derived from bone marrow supernatant (BM SN) from Ctrl mice (left) and ISO mice (right). DPP3 activity (U/L) in large (lEVs) and small (sEVs) EVs derived from (B) plasma of control mice (Ctrl) ( n = 6) and isoproterenol-treated mice (ISO) ( n = 5), or from (C) BM SN of Ctrl ( n = 6) and ISO mice ( n = 6). (D) DPP3 activity in lEVs and sEVs derived from BM SN of WT mice injected with bone marrow cells from Dpp3-KO (WT BM KO ) and Dpp3-KO mice injected with bone marrow cells from WT mice (KO BM WT ) ( n = 5) for each. (E) Quantification of Nano track analysis (NTA) results of plasma derived-lEVs and sEVs concentration (vesicle/ml) in healthy individuals ( n = 7), septic shock (SS) ( n = 6) and cardiogenic shock (CS) patients ( n = 5). (F) DPP3 activity (U/L) in lEVs and sEVs derived from plasma of healthy individuals ( n = 8) and septic shock (SS) patients ( n = 6). (G) DPP3 activity (U/L) in lEVs and sEVs derived from plasma of healthy individuals ( n = 8) and cardiogenic shock (CS) patients ( n = 11). DPP3 activity (U/L) in lEVs and sEVs derived from cell culture supernatant of (H) human cardiomyocytes AC16 ( n = 6) and (I) human monocytes THP-1 ( n = 2). In all bars, data are presented as mean ± standard error of the mean (SEM). Comparisons were made by Multiple Mann-Whitney tests in B, C, F, and G, and by two-way ANOVA in D and E. Significance was presented as follows (∗ p < 0.05, ∗∗ p < 0.01, and ns for non-significant differences).

Article Snippet: Three cell lines were used: human ventricular cardiomyocytes (AC16, Merck SCC109), murine ovarian cancer cells (ID8, Merck SCC145), and human monocytes (THP-1, kindly provided by Prof. Jean-Sébastien Silvestre).

Techniques: Western Blot, Derivative Assay, Activity Assay, Clinical Proteomics, Control, Injection, Concentration Assay, Cell Culture, MANN-WHITNEY

Journal: iScience

Article Title: Crosstalk between circulating DPP3 and immune cells in the context of cardio-systemic stress

doi: 10.1016/j.isci.2026.115114

Figure Lengend Snippet: A fraction of DPP3 is transported via EVs and upregulated during septic shock in humans (A) Western blot qualitative analysis of cluster of differentiation 63 (CD63) at 26 KD (bottom), heat shock protein 70 (HSC70) at 73 KD (middle), and DPP3 at 83 KD (above), in protein extracts of EVs derived from bone marrow supernatant (BM SN) from Ctrl mice (left) and ISO mice (right). DPP3 activity (U/L) in large (lEVs) and small (sEVs) EVs derived from (B) plasma of control mice (Ctrl) ( n = 6) and isoproterenol-treated mice (ISO) ( n = 5), or from (C) BM SN of Ctrl ( n = 6) and ISO mice ( n = 6). (D) DPP3 activity in lEVs and sEVs derived from BM SN of WT mice injected with bone marrow cells from Dpp3-KO (WT BM KO ) and Dpp3-KO mice injected with bone marrow cells from WT mice (KO BM WT ) ( n = 5) for each. (E) Quantification of Nano track analysis (NTA) results of plasma derived-lEVs and sEVs concentration (vesicle/ml) in healthy individuals ( n = 7), septic shock (SS) ( n = 6) and cardiogenic shock (CS) patients ( n = 5). (F) DPP3 activity (U/L) in lEVs and sEVs derived from plasma of healthy individuals ( n = 8) and septic shock (SS) patients ( n = 6). (G) DPP3 activity (U/L) in lEVs and sEVs derived from plasma of healthy individuals ( n = 8) and cardiogenic shock (CS) patients ( n = 11). DPP3 activity (U/L) in lEVs and sEVs derived from cell culture supernatant of (H) human cardiomyocytes AC16 ( n = 6) and (I) human monocytes THP-1 ( n = 2). In all bars, data are presented as mean ± standard error of the mean (SEM). Comparisons were made by Multiple Mann-Whitney tests in B, C, F, and G, and by two-way ANOVA in D and E. Significance was presented as follows (∗ p < 0.05, ∗∗ p < 0.01, and ns for non-significant differences).

Article Snippet: Human ventricular cardiomyocytes (AC16) , Merck , SCC109.

Techniques: Western Blot, Derivative Assay, Activity Assay, Clinical Proteomics, Control, Injection, Concentration Assay, Cell Culture, MANN-WHITNEY

Journal: Life Science Alliance

Article Title: RhoGEF Ect2 supports RhoA activity at cell–cell junctions through desmoplakin

doi: 10.26508/lsa.202503454

Figure Lengend Snippet: (A) Isolated cardiac myocytes (NRVCMs) were prepared for immunofluorescence and labeled for desmoplakin (DP), Ect2, and α-catenin and imaged using structured illumination microscopy. Orthogonal views can be found in . Scale bar = 5 μm. (B) Sections from control mouse hearts were fixed and stained for DP, Ect2, and N-cadherin (N-Cad) and imaged using the AxioVision Z1 system (Carl Zeiss) with Apotome slide module. Scale bar = 10 μm. (C) Proximity ligation assay (PLA) was performed on NRVCMs in control (Ctl KD) and DP knockdown (DP KD) conditions using antibodies directed against DP and Ect2. Scale bar = 20 μm. (C′) Quantification of each of three paired experiments was performed, showing significantly more PLA spots in Ctl KD conditions, indicating close proximity of DP and Ect2 (n = 3; PLA signal area for 17 control and 14 DP KD fields, paired t test,* P = 0.039). DAPI is used to stain nuclei in (A, B, C).

Article Snippet: Neonatal rat ventricular cardiomyocytes were isolated as described from 1- to 3-d-old Sprague-Dawley rats (Charles River) ( ).

Techniques: Isolation, Immunofluorescence, Labeling, Microscopy, Control, Staining, Proximity Ligation Assay, Knockdown

Journal: Life Science Alliance

Article Title: RhoGEF Ect2 supports RhoA activity at cell–cell junctions through desmoplakin

doi: 10.26508/lsa.202503454

Figure Lengend Snippet: (A) Isolated rat cardiac myocyte (NRVCM) structured illumination microscopy images with maximum image projections of single fluorophores and orthogonal planes illustrating the colocalization of Ect2 with junction proteins. Images are associated with the merged images in . Scale bar = 5 μm. (B, C) Sections from control mouse hearts (B) and control human hearts (C) were prepared for immunofluorescence and stained with antibodies directed against DP or Ect2 and imaged using the AxioVision Z1 system with Apotome slide module. Scale bar = 20 μm. (D, E) Immunoprecipitation of DP was performed on the HL-1 murine cardiomyocyte line (D) and isolated primary normal human keratinocytes (E), and lysates were probed for DP and Ect2. MW lane, molecular weight ladder.

Article Snippet: Neonatal rat ventricular cardiomyocytes were isolated as described from 1- to 3-d-old Sprague-Dawley rats (Charles River) ( ).

Techniques: Isolation, Microscopy, Control, Immunofluorescence, Staining, Immunoprecipitation, Molecular Weight

Journal: Life Science Alliance

Article Title: RhoGEF Ect2 supports RhoA activity at cell–cell junctions through desmoplakin

doi: 10.26508/lsa.202503454

Figure Lengend Snippet: (A) Isolated cardiac myocytes (NRVCMs) were treated with adenovirus encoding scrambled control (Ctl), DP, or Ect2 knockdown (KD) oligonucleotides, and then stained for DP and Ect2, as well as plakoglobin (Pg), to mark cell–cell borders. In addition, DP KD was rescued by expression of full-length desmoplakin II (DPII-FL). Yellow arrows indicate cell junctions containing DP and Ect2. (A′) Quantification of fluorescence intensity at the cell–cell junctions showed that Ect2 junctional staining was lost in the absence of DP and could be rescued by ectopic DPII expression (n = 3; ROIs: Ctl = 110, DP KD = 71, Ect2 KD = 67, and DP KD + DPII rescue = 58, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test, * P < 0.05). (B) NRVCMs were treated with adenovirus encoding scrambled control (Ctl) or DP KD oligonucleotides and stained for Ect2 and the adherens junction proteins N-cadherin (N-Cad) or α-catenin (α-Cat). (B′) Quantification of fluorescence intensity at the cell–cell junctions showed that neither N-cadherin nor α-catenin was significantly changed with DP KD, but Ect2 border intensity significantly decreased (n = 3; junction measurements: N-Cad/Ect2 = 44 Ctl, 62 DP KD; and α-catenin/Ect2 = 56 Ctl 68 DP KD, paired t test, P ≤ 0.05).

Article Snippet: Neonatal rat ventricular cardiomyocytes were isolated as described from 1- to 3-d-old Sprague-Dawley rats (Charles River) ( ).

Techniques: Isolation, Control, Knockdown, Staining, Expressing, Fluorescence

Journal: Life Science Alliance

Article Title: RhoGEF Ect2 supports RhoA activity at cell–cell junctions through desmoplakin

doi: 10.26508/lsa.202503454

Figure Lengend Snippet: (A) Isolated cardiac myocytes (NRVCMs) were treated with scrambled control (Ctl), DP, or Ect2 knockdown (KD) oligonucleotides, and then stained for plakoglobin (Pg) to mark cell–cell borders, as well as an antibody that recognizes active RhoA (Rho-GTP). Active RhoA cell junction localization is lost upon either DP or Ect2 silencing, and its border localization is restored by ectopic expression of DPII-FL. (A′) Quantification of Rho-GTP intensity in (A) (n = 3; ROIs: Ctl KD = 92 IDs, DP KD = 85 IDs, Ect2 KD = 80 IDs, DP KD + DPII-FL rescue = 70 IDs, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test). (B) Sections from control and Dsp cKO mouse hearts were fixed and triple-labeled for Pg to mark IDs, DP, and active RhoA. (B′) Quantification of DP border intensity in IDs of the WT versus Dsp cKO animals and Rho-GTP intensity at IDs of the WT and Dsp cKO animals showed a significant decrease in active RhoA at IDs with low DP localization compared with WT (DP low, * P = 0.011) or low DP localization IDs compared with IDs that retain DP (DP high IDs, * P = 0.009) (n = 3; 5–10 fields per condition per n with >10 IDs per field, paired t test for DP intensity, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test for RhoA intensity). Red, orange, and yellow insets present magnifications of IDs in the cardiac tissue. (C) Lysates from Ctl or DP KD HL-1 cardiac myocytes were subjected to RhoA (17A) pull-down to isolate active GEFs, and lysates were probed for Ect2. Whole-cell lysates (WCLs) were probed for DP and Ect2, as well as GAPDH as a loading control. (C′) Quantification of blots demonstrated a significant decrease in active Ect2 in DP KD lysates (n = 3; paired t test, P = 0.003). Scale bars = 20 μm.

Article Snippet: Neonatal rat ventricular cardiomyocytes were isolated as described from 1- to 3-d-old Sprague-Dawley rats (Charles River) ( ).

Techniques: Isolation, Control, Knockdown, Staining, Expressing, Labeling