Journal: Nature

Article Title: Nitrosative Stress Drives Heart Failure with Preserved Ejection Fraction

doi: 10.1038/s41586-019-1100-z

Figure Lengend Snippet: a, Left ventricular (LV) mRNA levels of Xbp1s, Bip, ATF6, ATF4, CHOP in mice of different experimental groups (n=5 mice per group). b, Immunoblot images of PERK, ATF6 and GAPDH proteins in LV samples of CHOW and HFD+L-NAME mice (n=3 mice per group). c, Electrophoretic analysis of spliced (s) and unspliced (u) Xbp1 transcript in LV samples of CHOW and HFD+L-NAME mice. Tunicamycin-treated neonatal rat ventricular myocytes (TUN) were used as positive control. Images are representative of three independently performed experiments with similar results. d, Adult mouse ventricular myocytes (AMVMs) mRNA levels of Xbp1s, Bip, CHOP in CHOW and HFD+L-NAME mice (n=4 mice per CHOW group; n=5 mice per HFD+L-NAME group. AMVMs were isolated from individual mice). e, Immunoblot images of LV pIRE1α, IRE1α and GAPDH proteins of CHOW and HFD+L-NAME mice (n=3 mice per group). f, Densitometric analysis ratio between pIRE1α and IRE1α protein bands (n=3 mice per group). g, mRNA levels of Xbp1s and Xbp1u in human myocardial biopsies from non-failing (CTR), HFpEF and HFrEF subjects (for CTR group n=15 subjects, for HFpEF group n=13 (Xbp1s) and n=14 (Xbp1u) subjects and for HFrEF group n=15). h, Immunoblot images of pIRE1α and IRE1α proteins in human myocardial biopsies from CTR, HFpEF and HFrEF subjects (n=8 subjects per group). i, Densitometric analysis ratio between pIRE1α and IRE1α protein bands (n=8 subjects per group). j, Ratio between mitral E wave and A wave (E/A) and k, Ratio between mitral E wave and E’ wave (E/E’) of control (CTR) and Xbp1s transgenic mice (TG) fed with CHOW or HFD+L-NAME diet over time (n=5 mice per CHOW CTR and CHOW TG groups; n=7 mice per HFD+L-NAME CTR and HFD+L-NAME TG groups. Each mouse was analyzed at all three time points). l, Running distance during exercise exhaustion test and m, Ratio between wet and dry lung weight (LW) at the end of the study (n=5 mice per CHOW CTR and CHOW TG groups; n=7 mice per HFD+L-NAME CTR and HFD+L-NAME TG groups). Results are presented as mean±S.E.M. a, g, i One-way ANOVA followed by Sidak’s multiple comparisons test. d, f, Two-tailed unpaired Student’s t-test. j-m Two-way ANOVA followed by Sidak’s multiple comparisons test. Numbers above square brackets show significant P values. For gel source data, see Supplementary Fig. 1.

Article Snippet: Human myocardial tissue Human myocardial tissue samples were collected under protocols approved by Institutional Review Board (IRB) at Johns Hopkins University (Maryland, USA) and consent for biopsy procedures or use of explanted tissues prospectively obtained in all cases.

Techniques: Western Blot, Positive Control, Isolation, Control, Transgenic Assay, Two Tailed Test

Journal: Nature

Article Title: Nitrosative Stress Drives Heart Failure with Preserved Ejection Fraction

doi: 10.1038/s41586-019-1100-z

Figure Lengend Snippet: a, Left ventricular (LV) mRNA levels of iNOS of different experimental groups of mice (n=4 mice per CHOW group; n=6 mice per HFD, L-NAME and HFD+L-NAME groups b, Adult mouse ventricular myocytes (AMVMs) mRNA levels of iNOS CHOW and HFD+L-NAME mice (n=5 mice per group. AMVMs were isolated from individual mice). c, LV mRNA levels of iNOS from WKY and ZSF1-Obese rats (n=5 rats per group). d, mRNA levels of iNOS in human myocardial biopsies from non-failing (CTR), HFpEF and HFrEF subjects (n=11 subjects per CTR group, n=11 subjects per HFpEF group and n=10 subjects per HFrEF group). e, Immunoblot images of S-nitrosylated IRE1α (SNO-IRE1α), IRE1α and GAPDH proteins in LV samples of wild type (WT) and iNOS knock out (iNOS KO) mice after five weeks of CHOW or HFD+L-NAME diet. Images are representative of four independently performed experiments with similar results. (- Asc: without Ascorbate, + Asc: with Ascorbate GSNO: S-Nitrosoglutathione) f, Densitometric analysis SNO-IRE1α +Ascorbate protein bands in each group (n=4 mice per group). g, Xbp1s mRNA level of neonatal rat ventricular myocytes transduced with increasing multiplicity of infection of of α-galactosidase adenovirus (AdLacZ) or iNOS adenovirus (AdiNOS) for 24 hours (n=4 biologically independent experiments). h, Immunoblot images of SNO-IRE1α and IRE1α proteins in neonatal rat ventricular myocytes transduced with adenovirus for IRE1α wild type (AdIRE1α WT) or AdIRE1α mutated in two target nitrosylation sites (AdIRE1α M1+M2) and AdlacZ or AdiNOS for 24 hours (- Asc: No Ascorbate. GSNO: S-Nitrosoglutathione). Images are representative of three independently performed experiments with similar results. i, Densitometric analysis ratio between SNO-IRE1α and IRE1α protein band intensities (n=3 biologically independent experiments). j, Xbp1s mRNA level of neonatal rat ventricular myocytes transduced with AdIRE1α WT or AdIRE1α M1+M2 and AdlacZ or AdiNOS for 24 hours (n=5 biologically independent experiments). Results are presented as mean±S.E.M. a, d, g, i, j One-way ANOVA followed by Sidak’s multiple comparisons test. b, c Two-tailed unpaired Student’s t-test. f, Two-way ANOVA followed by Sidak’s multiple comparisons test. Numbers above square brackets show significant P values. For gel source data, see Supplementary Fig. 1.

Article Snippet: Human myocardial tissue Human myocardial tissue samples were collected under protocols approved by Institutional Review Board (IRB) at Johns Hopkins University (Maryland, USA) and consent for biopsy procedures or use of explanted tissues prospectively obtained in all cases.

Techniques: Isolation, Western Blot, Knock-Out, Transduction, Infection, Two Tailed Test

Journal: Scientific Reports

Article Title: Disrupting the interaction between connexin 43 and calmodulin restores gap junction function and mitigates reperfusion arrhythmias

doi: 10.1038/s41598-025-18366-3

Figure Lengend Snippet: The establishment of isolated rat heart IR injury model. ( A ) Schematic representation of the experimental protocol of isolated rat heart IR injury model. ( B ) Representative TTC staining images showing myocardial infarct size after I/R injury. White areas indicate infarcted myocardium, while red areas represent viable (non-infarcted) myocardium. ( C ) Quantitative analysis of myocardial infarct size expressed as a percentage of the total myocardial area. ( D ) HE staining was performed to observe the I/R-induced histological changes. ( E ) Typical arrhythmias in the I/R group, the red arrows indicate premature electrical events, while the blue arrows indicate ST-segment elevation.

Article Snippet: Paraffin-embedded myocardial tissue Sect. (5 μm thick) were first deparaffinized in xylene (A530011, Sangon Biotech, China) and rehydrated through a series of graded ethanol solutions (A500737, Sangon Biotech, China).

Techniques: Isolation, Staining

Journal: Scientific Reports

Article Title: Disrupting the interaction between connexin 43 and calmodulin restores gap junction function and mitigates reperfusion arrhythmias

doi: 10.1038/s41598-025-18366-3

Figure Lengend Snippet: Interfering with the interaction between Cx43 and CaM improved myocardial electrophysiological parameters and reduced arrhythmia scores. (A) Recorded representative epicardial activation mapping of left ventricular. (B-D) Quantification of conduction velocity, absolute inhomogeneity, and inhomogeneity index of left ventricular. (E) Quantification of arrhythmia score in different groups. Data are expressed as the mean ± SD ( n = 6). Statistical differences were determined using one-way ANOVA, followed by post hoc Tukey’s test. *** P < 0.001 I/R group vs. control group; # P < 0.05 and ## P < 0.01 IR + sh-CaM#1 group vs. I/R group; & P < 0.05, && P < 0.01 and &&& P < 0.001 IR + SP15 group vs. I/R group.

Article Snippet: Paraffin-embedded myocardial tissue Sect. (5 μm thick) were first deparaffinized in xylene (A530011, Sangon Biotech, China) and rehydrated through a series of graded ethanol solutions (A500737, Sangon Biotech, China).

Techniques: Activation Assay, Control