Journal: Nature Communications

Article Title: Loss of TRIM29 mitigates viral myocarditis by attenuating PERK-driven ER stress response in male mice

doi: 10.1038/s41467-024-44745-x

Figure Lengend Snippet: a Survival of Trim29 fl/fl and cardiomyocyte-specific TRIM29 knockout ( Trim29 MyHC-KO ) mice after intraperitoneal infection with CVB3 (1 × 10 7 PFU per mouse) ( n = 10 per group). b Hematoxylin and eosin (H&E)-staining of heart sections from Trim29 fl/fl and Trim29 MyHC-KO mice after intraperitoneal infection without (Mock) or with CVB3 (1 × 10 7 PFU per mouse) for 4 days. Scale bars represent 1000 μm for original images and 400 µm for enlarged images. c Representative M-mode echocardiography images of hearts from Trim29 fl/fl and Trim29 MyHC-KO mice on day 4 after CVB3 infection. Cardiac function analysis of ejection fraction (EF) ( d ) and fractional shortening (FS) ( e ) of hearts from mice as in ( c ) ( n = 5 per group). f Assessment of heart weight/baseline body weight in Trim29 fl/fl and Trim29 MyHC-KO mice ( n = 5 per group) on day 0 or day 6 after CVB3 infection. ELISA of creatine kinase production in sera ( g ) and viral titers in homogenates of heart, pancreas, and spleen ( h ) from Trim29 fl/fl and Trim29 MyHC-KO mice on day 0 (Mock) and day 2 after CVB3 infection ( n = 5 per group). ELISA of IFN-α ( i ), IFN-β ( j ) IL-6 ( k ), TNF-α ( l ) and IL-1β ( m ) in hearts from Trim29 fl/fl and Trim29 MyHC-KO mice on day 2 after CVB3 infection ( n = 5 per group). Data are shown as the mean ± SD. Statistical significance was determined by a two-tailed, unpaired Student’s t test and Gehan-Breslow-Wilcoxon test for survival analysis. NS, not significant. Data are representative of three independent experiments. Source data are provided as a Source Data file.

Article Snippet: A mouse creatine kinase ELISA kit (NBP2-75306) was purchased from Novus Biologics.

Techniques: Knock-Out, Infection, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Loss of TRIM29 mitigates viral myocarditis by attenuating PERK-driven ER stress response in male mice

doi: 10.1038/s41467-024-44745-x

Figure Lengend Snippet: a Schematic illustration of the animal experiment. b Survival of wild-type (WT) mice after intraperitoneal infection with CVB3 (1 × 10 7 PFU per mouse) and treatment with the PERK inhibitor GSK2656157 or DMSO ( n = 10 per group). c Hematoxylin and eosin (H&E)-staining of heart sections from WT mice after intraperitoneal infection without (Mock) or with CVB3 (1 × 10 7 PFU per mouse) and treatment with the PERK inhibitor GSK2656157 or DMSO for 4 days. Scale bars represent 1000 μm for original images and 400 µm for enlarged images. d Representative M-mode images of hearts from WT mice after intraperitoneal infection without (Mock) or with CVB3 and treatment with the PERK inhibitor GSK2656157 or DMSO for 4 days by echocardiography analysis. Cardiac function analysis of ejection fraction (EF) ( e ) and fractional shortening (FS) ( f ) of hearts from mice as in ( d ) ( n = 5 per group). g Assessment of heart weight/baseline body weight in WT mice ( n = 5 per group) on day 0 or day 6 after CVB3 infection and treatment with the PERK inhibitor GSK2656157 or DMSO. ELISA of creatine kinase production in sera ( h ) and viral titers in homogenates of hearts ( i ) from WT mice on day 0 (Mock), day 2 and day 4 after CVB3 infection and treatment with the PERK inhibitor GSK2656157 or DMSO ( n = 5 per group). ELISA of IFN-α ( j ), IFN-β ( k ) IL-6 ( l ), TNF-α ( m ) and IL-1β ( n ) in hearts from WT mice on day 2 after CVB3 infection and treatment with the PERK inhibitor GSK2656157 or DMSO ( n = 5 per group). Data are shown as the mean ± SD. Statistical significance was determined by a two-tailed, unpaired Student’s t test and Gehan-Breslow-Wilcoxon test for survival analysis. NS, not significant. Data are representative of three independent experiments. Source data are provided as a Source Data file.

Article Snippet: A mouse creatine kinase ELISA kit (NBP2-75306) was purchased from Novus Biologics.

Techniques: Infection, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Science Advances

Article Title: Influenza virus replication in cardiomyocytes drives heart dysfunction and fibrosis

doi: 10.1126/sciadv.abm5371

Figure Lengend Snippet: WT and IFITM3 KO mice were intranasally infected with PR8-miR133b/206 or PR8-miRctrl (50 TCID 50 ). ( A ) Hearts were collected on day 10 after infection, and sections were stained with Masson’s trichrome stain, in which blue staining is indicative of fibrotic collagen deposition. Histological processing and image acquisition were performed by the OSU Comparative Pathology and Mouse Phenotyping Core Facility on heart tissue samples provided by A.D.K. A representative heart section is shown for each genotype-virus combination. Boxed areas are regions magnified in the far-right images. Scale bars, 1 mm and 200 μm for the left and right images, respectively. ( B ) Percent fibrosis was calculated by quantifying ratio of blue pixel intensity to total pixel intensity for each heart section. Each point represents a heart from an individual mouse, and bars represent mean values. Error bars represent SD of the mean. Comparisons were analyzed by ANOVA followed by Tukey’s post hoc test. * P < 0.05. ( C ) Serum from IFITM3 KO mice was collected before infection and at days 5 and 10 after infection with PR8-miRctrl or PR8-miR133b/206 for ELISA quantification of creatine kinase. Data points represent individual mice, and bars represent mean values. Error bars depict SD of the mean. Comparisons were analyzed by ANOVA followed by Tukey’s post hoc test. * P < 0.05.

Article Snippet: Creatine kinase quantification was performed using the Mouse Creatine Kinase MB ELISA Kit (Novus Biologicals).

Techniques: Infection, Staining, Virus, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: The long noncoding RNA lncCIRBIL disrupts the nuclear translocation of Bclaf1 alleviating cardiac ischemia–reperfusion injury

doi: 10.1038/s41467-020-20844-3

Figure Lengend Snippet: A Bclaf1 protein level determined by western blot. N = 3. * P < 0.05 versus WT group. P values were determined by unpaired t test. B Representative images of echocardiographs and statistics of ejection fraction (EF) and fractional shortening (FS). WT N = 13, Bclaf1-TG N = 11, WT-I/R N = 10, Bclaf1-TG-I/R N = 13. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. C Infarct area determined by TTC staining (scale bar: 2 mm). N = 6 mice per group. * P < 0.05 versus WT-IR mice. P values were determined by unpaired t test. D , E Serum concentrations of LDH and CKMB, respectively, measured by Elisa assay. N = 9 mice per group. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. F , G Caspase-3 activity and cleaved caspase-3 protein level, respectively. N = 8 mice per group for caspase activity, and N = 3 mice for cleaved caspase-3 protein level. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. H Apoptosis of cardiomyocyte in the border zone (BZ) by TUNEL staining (scale bar: 20 μm). N = 7 mice per group. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. I mRNA and protein levels of p53 by RT-PCR and western blot. N = 9 for PCR, N = 3 for western blot. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. J mRNA and protein levels of Bax by RT-PCR and western blot. N = 9 for PCR, N = 4 for western blot. * P < 0.05 versus WT, # P < 0.05 versus WT-IR mice. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. K Distribution of Bclaf1 in isolated cardiomyocytes (scale bar: 20 μm). Data are expressed as mean ± SEM.

Article Snippet: The CKMB was detected by using the mouse CKMB (Creatine Kinase MB Isownzyme) Elisa Kit (Elabscience, Wuhan, China).

Techniques: Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, TUNEL Assay, Reverse Transcription Polymerase Chain Reaction, Isolation

Journal: Nature Communications

Article Title: The long noncoding RNA lncCIRBIL disrupts the nuclear translocation of Bclaf1 alleviating cardiac ischemia–reperfusion injury

doi: 10.1038/s41467-020-20844-3

Figure Lengend Snippet: A Bclaf1 in the heart tissue after adeno-associated virus carrying cas9-Bclaf1 injection. N = 6. * P < 0.05 versus NC group. P values were determined by unpaired t test. B Representative images of echocardiographs and statistics of ejection fraction (EF) and fractional shortening (FS). NC N = 11, Bclaf1-pKO N = 9, NC-I/R N = 13, Bclaf1-pKO-I/R N = 18. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. C Infarct size by TTC staining (scale bar: 2 mm). NC-I/R N = 5, Bclaf1-pKO-I/R N = 10. * P < 0.05 versus NC-I/R, P values were determined by unpaired t test. D , E Serum concentrations of LDH and CKMB, respectively, measured by Elisa assay. N = 9 mice per group. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. F Caspase-3 activity. N = 8 mice per group. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. G Cleaved caspase-3 protein level. N = 3 mice per group. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. H Apoptosis of cardiomyocyte in the border zone (BZ) by TUNEL staining (scale bar: 20 μm). N = 8 mice per group. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. I The mRNA and protein levels of p53, respectively. N = 9 for mRNA and N = 3 for protein. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. J The mRNA and protein levels of Bax, respectively. N = 9 for mRNA and N = 4 for protein. * P < 0.05 versus NC, # P < 0.05 NC-IR. P values were determined by one-way ANOVA test followed by Bonferroni post hoc analysis. Data are expressed as mean ± SEM.

Article Snippet: The CKMB was detected by using the mouse CKMB (Creatine Kinase MB Isownzyme) Elisa Kit (Elabscience, Wuhan, China).

Techniques: Virus, Injection, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, TUNEL Assay