Journal: Foods

Article Title: Sea Buckthorn Pericarp Flavonoids Improve Diet-Induced Hyperlipidemia via Coordinated Modulation of Hepatic Lipid Metabolism and Gut Microbiota

doi: 10.3390/foods15061049

Figure Lengend Snippet: TFSP modulates the expression of ACC, FAS, CPT-1α, PPARα and ATGL proteins in the livers of HFD mice. n = 6 per group. Data are presented as mean ± SD. Different letters above bars indicate statistically significant differences ( p < 0.05) by one-way ANOVA followed by LSD post hoc test.

Article Snippet: Peroxisome proliferator-activated receptor alpha (PPARα) primary antibody (Cat. No. A00600-2), carnitine palmitoyltransferase-1 alpha (CPT-1α) primary antibody (Cat. No. A00917-3), acetyl-CoA carboxylase (ACC) primary antibody (Cat. No. M01802-2), fatty acid synthase (FAS) primary antibody (Cat. No. BA0484), adipose triglyceride lipase (ATGL) primary antibody (Cat. No. A01800-1), and GAPDH primary antibody (Cat. No. BM1623) were obtained from BOSTER Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 is upregulated in heart tissues of mice challenged with Ang II. (A) RNA sequencing was performed on mouse cardiac tissues to analyze the expression profile of E3 ubiquitin ligases (red indicates up‐regulated genes, green indicates down‐regulated genes). Gray dots represent E3 ubiquitin ligases with no statistically significant difference compared to the control group. A fold change > 2 and p < 0.05 were considered statistically significant. (B) Representative western blot analysis of TRIM40 protein levels in cardiac tissues from Ctrl and Ang II‐induced mice. Glyceraldehyde‐3 phosphate dehydrogenase (GAPDH) was used as a loading control (n = 6). (C) Representative western blot analysis of TRIM40 protein levels in cardiac tissues from Ctrl and transverse aortic constriction (TAC)‐induced mice. GAPDH was used as a loading control (n = 6). (D) Immunofluorescence staining of mouse cardiac tissues showing the localization of TRIM40 (red), vimentin (green), and sarcomeric α‐actinin (green). Nuclei were counterstained with 4’,6‐Diamidino‐2’‐phenylindole (DAPI) (blue). Scale bar = 50 µm (n = 6). (E) Representative western blot analysis of TRIM40 protein levels in macrophages, NRVMs, NRCFs, and H9c2 cells. GAPDH was used as a loading control (n = 3).

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: RNA Sequencing, Expressing, Ubiquitin Proteomics, Control, Western Blot, Immunofluorescence, Staining

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 deficiency protects against Ang II‐induced myocardial hypertrophy and fibrosis. WT and TRIM40 −/− mice were infused with saline or Ang II for 4 weeks. (A) Mouse systolic blood pressure was measured weekly using a non‐invasive tail‐cuff method (n = 6). (B) Serum Ang II concentration was detected in mice (n = 6). (C) Representative echocardiographic images of mice from each group (n = 6). (D–F) Cardiac function parameters showing EF (D), FS (E), and IVRT (F) (n = 6). (G) Serum CK‐MB levels were measured using an ELISA kit (n = 6). (H) HW/BW of mice in each group (n = 6). (I) Representative freshly isolated heart specimens photographed against a white background (scale bar = 5 mm) (n = 6). (J) Cardiomyocyte cross‐sectional area was assessed by dual immunofluorescence staining for cTnT (red) to mark cardiomyocytes and fluorescein‐labeled WGA (green) to delineate cell membranes (scale bar = 50 µm) (n = 6). (K) H&E staining of heart tissue sections (scale bar = 50 µm) (n = 6). (L, M) Myocardial fibrosis was evaluated by Masson's trichrome staining (L) and Picrosirius red staining (M) (scale bar = 50 µm) (n = 6). (N) Representative western blot analyses of MYH7, ANP, and BNP in heart tissue, with GAPDH as a loading control (n = 6). (O) Densitometric quantification of the blots in N (n = 6). (P, Q) mRNA expression levels of hypertrophy‐associated genes (P) and inflammation‐related genes (Q) in heart tissues of mice. Data was normalized to Actb (n = 6). All quantitative data are presented as mean ± SEM. Data between two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test. ns indicates not statistically significant; * p < 0.05, ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Saline, Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation, Immunofluorescence, Staining, Labeling, Western Blot, Control, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 deficiency prevents TAC‐induced myocardial hypertrophy and fibrosis. WT and TRIM40 −/− mice were subjected to TAC to model pressure overload‐induced cardiac hypertrophy and remodeling. (A) Representative echocardiographic images of mice from each group (n = 6). (B–D) Cardiac function parameters showing EF (B), FS (C), and IVRT (D) (n = 6). (E) Serum CK‐MB levels were measured using an ELISA kit (n = 6). (F) HW/BW of mice in each group (n = 6). (G) Representative freshly isolated heart specimens photographed against a white background (scale bar = 5 mm) (n = 6). (H) Cardiomyocyte cross‐sectional area was assessed by dual immunofluorescence staining for cTnT (red) to mark cardiomyocytes and fluorescein‐labeled WGA (green) to delineate cell membranes (scale bar = 50 µm) (n = 6). (I) Quantitative analysis of cardiomyocyte cross‐sectional area. A minimum of 100 cells were measured from different fields across at least four samples per group (n = 6). (J) H&E staining of heart tissue sections (scale bar = 50 µm) (n = 6). (K, L) Myocardial fibrosis was evaluated by Masson's trichrome staining (K) and Picrosirius red staining (L) (scale bar = 50 µm) (n = 6). (M) Representative western blot analyses of MYH7, ANP, and BNP in heart tissue, with GAPDH as a loading control (n = 6). (N) Densitometric quantification of the blots in M (n = 6). (O, P) mRNA expression levels of hypertrophy‐associated genes (O) and inflammation‐related genes (P) in heart tissues. Data were normalized to Actb (n = 6). All quantitative data are presented as mean ± SEM. Data between two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test; ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Immunofluorescence, Staining, Labeling, Western Blot, Control, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: Cardiomyocyte‐specific knockdown of TRIM40 alleviates Ang II‐induced cardiac remodeling. (A) Mouse systolic blood pressure was measured weekly using a non‐invasive tail‐cuff method (n = 6). (B) Serum Ang II concentration was detected in mice (n = 6). (C) Representative echocardiographic images from each group of mice (n = 6). (D‐F) Cardiac function parameters: EF (D), FS (E), and IVRT (F) (n = 6). (G) Serum CK‐MB levels (n = 6). (H) Heart weight/body weight ratio (n = 6). (I) Representative images of freshly isolated heart specimens (n = 6). (J) Quantitative analysis of the cross‐sectional area of heart specimens shown in (I) (n = 6). (K) Cardiomyocyte cross‐sectional area was assessed by dual immunofluorescence staining for cTnT (red) to mark cardiomyocytes and fluorescein‐labeled WGA (green) to delineate cell membranes (scale bar = 50 µm) (n = 6). (L) Quantitative analysis of cardiomyocyte cross‐sectional area based on K. A minimum of 100 cells were measured from different fields across at least four samples per group (n = 6). (M) H&E staining of heart tissue sections (scale bar = 50 µm) (n = 6). (N, O) Myocardial fibrosis evaluated by Picrosirius Red (N) and Masson's trichrome (O) staining (scale bar = 50 µm) (n = 6). (P) Representative Western blots of MYH7, ANP, and BNP protein expression in heart tissue, with GAPDH as a loading control (n = 6). (Q) Densitometric quantification of the blots shown in (P) (n = 6). (R, S) mRNA expression levels of hypertrophy‐associated (R) and inflammation‐related (S) genes in heart tissues (n = 6). Data were normalized to Actb (n = 6). All quantitative data are presented as mean ± SEM. Data between the two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test. ns indicates not statistically significant; ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Knockdown, Concentration Assay, Isolation, Immunofluorescence, Staining, Labeling, Western Blot, Expressing, Control, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 regulates cardiomyocyte hypertrophic and fibrotic responses in culture. (A) After transfection with either a negative control (NC) or TRIM40 siRNA (si‐TRIM40), NRVMs were treated with Ang II (1 µ m for 24 h). Cardiomyocyte size was assessed by dual immunofluorescence staining for cTnT (green) and Rhodamine‐phalloidin (red). Nuclei were counterstained with DAPI (blue) (scale bar = 50 µm) (n = 3). (B) Changes in NRVMs size in response to Ang II. At least 100 cells from each of three independent samples were analyzed per group (n = 3). (C) Representative western blot of MYH7, ANP, and BNP proteins in NRVMs, with GAPDH as loading control (n = 3). (D, E) The mRNA levels of hypertrophy‐associated genes (D) and inflammation‐related genes (E) in NRVMs treated as indicated. Data was normalized to Actb (n = 3). (F) Following transfection with Flag‐TRIM40 or an empty vector (EV), NRVMs were treated with Ang II (1 µ m for 24 h). Cardiomyocyte size was assessed by the dual immunofluorescence staining method described in (A) (scale bar = 50 µm) (n = 3). (G) Changes in NRVMs size in response to Ang II. At least 100 cells from each of three independent samples were analyzed per group (n = 3). (H) Representative western blot of MYH7, ANP, and BNP proteins in NRVMs, with GAPDH as loading control (n = 3). (I, J) mRNA expression levels of hypertrophy‐associated genes (I) and inflammation‐related genes (J) in NRVMs. Data were normalized to Actb (n = 3). All quantitative data are presented as mean ± SEM. Data between the two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test; * p < 0.05, ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Transfection, Negative Control, Immunofluorescence, Staining, Western Blot, Control, Plasmid Preparation, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 interacts directly with PKN2. (A) Schematic diagram of the quantitative proteomic screening workflow for identifying TRIM40‐interacting proteins. (B) Tandem mass spectrum of representative peptide fragments from PKN2. (C) Amino acid sequence information of the identified PKN2 peptides. (D, E) Co‐IP assays using anti‐Flag antibody in NRVMs (D) and HEK‐293T cells (E) transfected with Flag‐tagged TRIM40, followed by immunoblotting to detect PKN2 association. IgG served as a NC for Co‐IP (n = 3). (F) Endogenous PKN2 binding was detected by immunoblotting after Co‐IP with anti‐TRIM40 antibody from mouse heart tissue lysates. IgG was used as a control (n = 6). (G) Schematic representation of the domain deletion mutants of PKN2. (H) HEK‐293T cells were co‐transfected with HA‐tagged full‐length PKN2 or its deletion mutants together with Flag‐TRIM40. Immunoprecipitation was performed using anti‐HA antibody, followed by immunoblotting to detect Flag‐TRIM40 binding (n = 3). (I) Schematic diagrams of TRIM40 domain deletion mutants and its catalytically inactive mutant (C29S). (J) HEK‐293T cells were co‐transfected with Flag‐tagged full‐length TRIM40 or its mutants together with HA‐PKN2. Immunoprecipitation with anti‐Flag antibody was used to assess HA‐PKN2 binding (n = 3). (K) Ubiquitination assay of PKN2 in HEK‐293T cells co‐expressing Myc‐Ub, HA‐PKN2, and the catalytically inactive mutant Flag‐TRIM40‐C29S. HA immunoprecipitates were analyzed by immunoblotting to detect PKN2 ubiquitination. (n = 3) (L) Confocal microscopy images showing the effects of different TRIM40 variants (full‐length, deletion mutants, and C29S mutant) on F‐actin cytoskeleton organization in NRVMs. Cells were stained with rhodamine‐conjugated phalloidin (red, labeling F‐actin) and anti‐TRIM40 antibody (green, indicating transfected TRIM40 variants), with nuclei counterstained by DAPI (blue). Scale bar = 50 µm (n = 3). (M) Structural basis of the TRIM40‐PKN2 interaction.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Sequencing, Co-Immunoprecipitation Assay, Transfection, Western Blot, Binding Assay, Control, Immunoprecipitation, Mutagenesis, Ubiquitin Proteomics, Expressing, Confocal Microscopy, Staining, Labeling

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 Positively Regulates the Phosphorylation of PKN2 by Mediating Its K63‐Linked Ubiquitination. (A) HEK‐293T cells were transfected with HA‐PKN2, Myc‐Ub, and Flag‐TRIM40, and treated with 10 µ m MG132 for 6 h before harvesting. The ubiquitination level of PKN2 was detected by immunoblotting using an anti‐HA antibody (Control = IgG) (n = 3). (B) HEK‐293T cells were co‐transfected with HA‐PKN2, Flag‐TRIM40, and various types of Myc‐Ub (including WT, K6‐, K11‐, K27‐, K29‐, K33‐, K48‐, and K63‐linked ubiquitin chains). After immunoprecipitation with anti‐HA magnetic beads, the ubiquitination of PKN2 was analyzed by immunoblotting. Cells were pretreated with 10 µ m MG132 for 6 h before harvesting (n = 3). (C) HEK‐293T cells were transfected with HA‐PKN2, Myc‐Ub, EV, Flag‐TRIM40, and Flag‐TRIM40‐29S, treated with 10 µ m MG132 for 6 h before harvesting. The ubiquitination level of PKN2 was detected by immunoblotting using an anti‐HA antibody (n = 3). (D) NRVMs were transfected with siRNA targeting TRIM40, followed by treatment with 1 µ m Ang II for 12 h. The phosphorylation level of PKN2 was detected by immunoblotting, with GAPDH used as the loading control (n = 3). (E) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel D (n = 3). (F) NRVMs were transfected with a TRIM40 expression vector, followed by treatment with 1 µ m Ang II for 12 h. The phosphorylation level of PKN2 was detected by immunoblotting, with GAPDH used as the loading control (n = 3). (G) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel F (n = 3). (H) NRVMs overexpressing Flag‐TRIM40 or Flag‐TRIM40‐C29S were treated with Ang II (1 µ m ) for 24 h. Phosphorylation of PKN2 at Ser815 (p‑PKN2) was detected by Western blot. GAPDH served as a loading control (n = 3). (I) Densitometric quantification of the Western blot bands from Figure (n = 3). (J) The level of p‐PKN2 was detected by immunoblotting in whole heart tissue lysates from mice infused with Ang II for 4 weeks (n = 6). (K) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel J (n = 6). (L) The level of p‐PKN2 was detected by immunoblotting in whole heart tissue lysates from mice subjected to TAC surgery (n = 6). (M) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel L (n = 6). (N) NRVMs overexpressing Flag‐TRIM40 or Flag‐TRIM40‐C29S were treated with Ang II (1 µ m ) for 24 h. Protein levels of MYH7, ANP and BNP were detected by Western blot. GAPDH served as a loading control (n = 3). (O) Densitometric quantification of the Western blot bands from Figure (n = 3). All quantitative data are presented as mean ± SEM. Data between two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test; ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Phospho-proteomics, Ubiquitin Proteomics, Transfection, Western Blot, Control, Immunoprecipitation, Magnetic Beads, Software, Expressing, Plasmid Preparation, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 exacerbates Ang II‐induced cardiomyocyte hypertrophy and fibrosis by regulating PKN2. C57BL/6 mice received two injections of AAV9 encoding TRIM40 at one‐month intervals, followed by continuous infusion of saline or Ang II for two weeks. (A) Mouse systolic blood pressure was measured weekly using a non‐invasive tail‐cuff system (n = 6). (B) Serum Ang II concentration was detected in mice (n = 6). (C) Representative echocardiographic images of mice from each experimental group (n = 6). (D–F) Cardiac function parameters showing EF (D), FS (E), and IVRT (F) (n = 6). (G) Serum CK‐MB levels in mice (n = 6). (H) HW/BW of mice in each group (n = 6). (I) Representative freshly isolated heart specimens photographed against a white background (scale bar = 5 mm) (n = 6). (J) Cardiomyocyte size was assessed by fluorescein‐conjugated WGA staining (scale bar = 50 µm) (n = 6). (K) H&E staining of heart tissue sections (scale bar = 50 µm) (n = 6). (L, M) Myocardial fibrosis was evaluated by Masson's trichrome staining (L) and Picrosirius red staining (M) (scale bar = 50 µm) (n = 6). (N) Western blot analysis of MYH7, ANP, and BNP in myocardial tissues, with GAPDH as a loading control (n = 6). (O, P) mRNA expression levels of hypertrophy‐associated genes (O) and inflammation‐related genes (P) in myocardial tissues, normalized to Actb (n = 6). (Q) p‐PKN2 levels were detected by immunoblotting in whole heart lysates from mice infused with Ang II for 4 weeks (n = 6). (R) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel Q (n = 6). All quantitative data are presented as mean ± SEM. Data between the two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by a Tukey post hoc test. ns indicates not statistically significant; * p < 0.05, ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Saline, Concentration Assay, Isolation, Staining, Western Blot, Control, Expressing, Software, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: TRIM40 exacerbates TAC‐induced cardiomyocyte hypertrophy and fibrosis by regulating PKN2. C57BL/6 mice received two injections of AAV9 encoding TRIM40 at one‐month intervals, followed by sham surgery or TAC surgery for two weeks. (A) Representative echocardiographic images of mice from each experimental group (n = 6). (B–D) Cardiac function parameters showing EF (B), FS (C), and IVRT (D) (n = 6). (E) Serum CK‐MB levels in mice (n = 6). (F) HW/BW of mice in each group (n = 6). (G) Representative freshly isolated heart specimens photographed against a white background (scale bar = 5 mm) (n = 6). (H) Cardiomyocyte size was assessed by fluorescein‐conjugated WGA staining (scale bar = 50 µm) (n = 6). (I) Cardiomyocyte hypertrophy evaluated by fluorescein‐conjugated WGA staining (scale bar = 50 µm) (n = 6). (J) H&E staining of heart tissue sections (scale bar = 50 µm) (n = 6). (K, L) Myocardial fibrosis was evaluated by Masson's trichrome staining (K) and Picrosirius red staining (L) (scale bar = 50 µm) (n = 6). (M) Western blot analysis of MYH7, ANP, and BNP in myocardial tissues, with GAPDH as a loading control (n = 6). (N, O) mRNA expression levels of hypertrophy‐associated genes (N) and inflammation‐related genes (O) in myocardial tissues, normalized to Actb (n = 6). (P) p‐PKN2 levels were detected by immunoblotting in whole heart lysates from mice infused with Ang II for 4 weeks (n = 6). (Q) Quantitative data of the blot intensity of corresponding proteins determined by Image J software in panel P (n = 6). All quantitative data are presented as mean ± SEM. Data between the two groups were compared by independent‐sample two‐tailed Student's t‐test. Data among multiple groups were compared by one‐way ANOVA test, followed by Tukey post hoc test; ** p < 0.01.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Isolation, Staining, Western Blot, Control, Expressing, Software, Two Tailed Test

Journal: Advanced Science

Article Title: TRIM40 Drives Pathological Cardiac Hypertrophy and Heart Failure via Ubiquitination of PKN2

doi: 10.1002/advs.202521337

Figure Lengend Snippet: A Mechanism of TRIM40 Driving Cardiac Hypertrophy through PKN2 Ubiquitination.

Article Snippet: Antibodies against TRIM40 (24526‐1‐AP), MYH7 (22280‐1‐AP), ANP (27426‐1‐AP), BNP (13299‐1‐AP), PKN2 (14608‐1‐AP), cTnT (68300‐1‐Ig), Rabbit IgG (B900610), His (6005‐1‐lg), Flag (20543‐1‐AP) and HA (51064‐2‐AP) were purchased from Proteintech (IL, USA).

Techniques: Ubiquitin Proteomics