trpc6 antibody  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Cardiac systolic and diastolic functions assessed by echocardiography in WT and TRPC6 KO. ( A ) Stroke volume, ( B ) cardiac output, ( C ) ejection fraction, ( D ) fractional shortening, ( E ) isovolumic relaxation time, ( F ) myocardial performance index, and ( G ) E/e’, measurements at baseline, 4 weeks, and 8 weeks after L-NAME. Results are expressed as mean ± SEM, n = 6–7, * p < 0.05 between two groups by post hoc test after two-way ANOVA. Bar colors indicate experimental groups: white = WT RD (regular diet), gray = WT HFD+L-NAME, light blue = KO RD, dark blue = KO HFD+L-NAME.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Left ventricle wall thickness and fibrosis in WT and TRPC6 KO mice. ( A ) End-systolic left ventricular anterior wall thickness, ( B ) end-diastolic left ventricular anterior wall thickness, ( C ) end-systolic left ventricular posterior wall thickness, and ( D ) end-diastolic left ventricular posterior wall thickness, with measurements at baseline, 4 weeks and 8 weeks after L-NAME. Results are expressed as mean ± SEM, n = 6–8, * p < 0.05 between two groups by post hoc test after two-way ANOVA. Myocardial fibrosis measurements in WT and TRPC6 KO mice. Representative Masson Trichrome staining of left ventricles from WT RD ( E ), WT HFD+ L-NAME ( F ), KO RD ( G ), and KO HFD+ L-NAME ( H ). ( I ) Quantitative analysis of collagen fractions (blue color) by Masson Trichrome staining in the left ventricles. Results are expressed as mean ± SEM, n = 6, * p < 0.05 between two groups by non-parametric test after one-way ANOVA. ns, not significant. Bar colors indicate experimental groups: white = WT RD (regular diet), gray = WT HFD+L-NAME, light blue = KO RD, dark blue = KO HFD+L-NAME.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Echocardiography during dobutamine stress test. Time-dependent changes after dobutamine injection in ( A ) Heart rate, ( B ) ejection fraction, ( C ) stroke volume, ( D ) cardiac output, ( E ) LVESV, and ( F ) LVEDV in WT and TRPC6 KO mice with RD. ( G – L ) Heart rate, ejection fraction, stroke volume, cardiac output, LVESV, and LVEDV in WT and TRPC6 KO mice with HFD+L-NAME after dobutamine injection. Results are expressed as mean ± SEM, n = 5–6, *, p < 0.05 when compared with baseline in WT mice, #, p < 0.05 when compared to baseline in TRPC6 KO mice by post hoc test after one-way ANOVA. LVESV, left ventricular end-systolic volume; LVEDV, left ventricular end-diastolic volume. Color code: for WT RD and WT HFD+L-NAME mice, white (baseline), light gray (1 min), medium gray (5 min), dark gray (10 min) after dobutamine injection; for KO-RD and KO HFD+L-NAME mice, light blue (baseline), medium blue (1 min), darker blue (5 min), dark blue (10 min) after dobutamine injection.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques: Injection

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Changes in cardiac function at 10 min after dobutamine compared to baseline. Changes in ( A ) heart rate, ( B ) ejection fraction, ( C ) stroke volume, ( D ) cardiac output, ( E ) LVESV, and ( F ) LVEDV in WT and TRPC6 KO mice with RD or HFD+ LNAME. Results are expressed as mean ± SEM, n = 5–6, * p < 0.05 between two groups by post hoc test after one-way ANOVA.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Treadmill running test in WT and TRPC6 KO mice. ( A ) Running time, ( B ) running distance, and ( C ) vertical work in WT and TRPC6 KO mice with RD or HFD+ L-NAME. Results are expressed as mean ± SEM, n = 5, * p < 0.05 between two groups by post hoc test after one-way ANOVA. Bar colors indicate experimental groups: white = WT RD (regular diet), gray = WT HFD+L-NAME, light blue = KO RD, dark blue = KO HFD+L-NAME.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Changes in protein expression within the cytosol and mitochondria isolated from the left ventricle of WT and TRPC6 KO mice. ( A ) Representative blots for caspase 9, cleaved caspase 9, PGC1a, TRPC6, TRPC3, BNP, and GAPDH in the cytosol fraction. ( B ) Quantitative analysis results for protein expression levels in the cytosol. ( C ) Representative blots for TRPC6, TRPC3, and VDAC in mitochondria isolation. ( D ) Quantitative analysis results for protein expression levels in mitochondria. Results are expressed as mean ± SEM, n = 3–6, * p < 0.05 between two groups by post hoc test after one-way ANOVA. Bar colors indicate experimental groups: white = WT RD (regular diet), gray = WT HFD+L-NAME, light blue = KO RD, dark blue = KO HFD+L-NAME.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques: Expressing, Isolation

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Immunohistochemistry staining of BNP and cleaved caspase 9 from the left ventricle of WT and TRPC6 KO mice with RD or HFD+ L-NAME. Representative images for BNP staining in WT RD ( A , E ), WT HFD+ L-NAME ( B , F ), KO RD ( C , G ), and KO HFD+ L-NAME ( D , H ), and cleaved caspase 9 staining in WT RD ( I , M ), WT HFD+ L-NAME ( J , N ), KO RD ( K , O ), and KO HFD+ L-NAME ( L , P ). Upper panel, ×40 magnification; lower panel, boxed regions in ×200 magnification.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques: Immunohistochemistry, Staining

Journal: International Journal of Molecular Sciences

Article Title: TRPC6 Deficiency Attenuates Mitochondrial and Cardiac Dysfunction in Heart Failure with Preserved Ejection Fraction Induced by High-Fat Diet Plus L-NAME

doi: 10.3390/ijms26199383

Figure Lengend Snippet: Mitochondrial respiration, mitochondria-derived H 2 O 2 production, and mitochondrial DNA copy numbers in WT and TRPC6 KO mice. ( A ) Maximal oxygen consumption rate, ( B ) ATP-linked oxygen consumption rate, ( C ) basal H 2 O 2 generation rate, and ( D ) maximal H 2 O 2 generation rate measurements by Oroboros respirometer in WT and TRPC6 KO mice with RD or HFD+ L-NAME. ( E , F ) Mitochondrial DNA copy number normalized to 18s rRNA and lipoprotein lipase, respectively. Results are expressed as mean ± SEM, n = 6, * p < 0.05 between two groups by post hoc test after one-way ANOVA. Bar colors indicate experimental groups: white = WT RD (regular diet), gray = WT HFD+L-NAME, light blue = KO RD, dark blue = KO HFD+L-NAME.

Article Snippet: After transfer to nitrocellulose membranes, blots were rinsed in PBS and blocked in Odyssey blocking buffer (LI-CORbio, Lincoln, NE, USA) for 1 h at room temperature and then incubated with TRPC6 rabbit antibody (1:500, ACC-120, Alomone Labs, Jerusalem, Israel) or TRPC3 rabbit antibody (1:500, #77934, Cell Signaling Technology, Danvers, MA, USA), VDAC rabbit antibody (1:1000, #4661, Cell Signaling), GAPDH rabbit antibody (1:2000, #2118, Cell Signaling), caspase 9 rabbit antibody (1:1000, #9504, Cell Signaling), cleaved caspase 9 rabbit antibody (1:1000, #7237, Cell Signaling), PGC-1α rabbit antibody (1:1000, #2178, Cell Signaling) and BNP rabbit antibody (1:1000, PA5-96084, ThermoFisher, Waltham, MA, USA) at 4 °C overnight.

Techniques: Derivative Assay

Journal: Advanced Science

Article Title: TBC1D22B Regulates ER‐to‐Golgi Trafficking via RAB1B Inactivation and Promotes Oncogenic Programs in Breast Cancer

doi: 10.1002/advs.202502269

Figure Lengend Snippet: Overexpression of TBC1D22B, but not of TBC1D22A, induces tumor spheroids overgrowth, in a GAP‐dependent manner. BT549 cells stably expressing the empty vector (EV), HA‐tagged TBC1D22B, RQ or TBC1D22A were grown in matrigel for 7 days to form tumor cell spheroids. Spheroids were stained with Phalloidin and DAPI shown respectively in magenta and grey in the merged representative images (left). Images are confocal Z‐stack projections (8 optical sections/image) of spheroids acquired with a 40x objective, zoom 1.5x. Bar, 50 µm. Quantification of the spheroids area, shown in the middle panel, is from 3 independent experiments in which 45–71 spheroids/condition were measured. Mean ± SD are also shown in the plots. Statistical analysis was performed with the t‐test, **** p < 0.0001. The Western blots on the right were IB with the indicated antibodies (vinculin, loading control) and show the expression levels of TBC1D22B, RQ and TBC1D22A. B) CAL120 cells stably expressing the empty vector (EV), the HA‐tagged TBC1D22B, or RQ were grown and stained as in A. Images are confocal Z‐stack projections (8 optical sections/image) of representative spheroids acquired with a 20x objective, zoom 2. Bar, 50 µm. In the middle panel the area of 45 spheroids/condition from 3 independent experiments is shown. Mean ± SD are also shown in the plots. Statistical analysis was performed with the t‐test, p values: ***, p < 0.001, **** p < 0.0001. The Western blot on the right shows the expression levels of TBC1D22B and RQ.

Article Snippet: CAL120 cells, from DSMZ, were grown in DMEM (ECM0749L, Euroclone) supplemented with 10% fetal bovine serum (ECS1800D, Euroclone), 1% L‐glutamine (ECB3000D, Euroclone) and 1% penicillin‐streptomycin (ECB3001D, Euroclone).

Techniques: Over Expression, Stable Transfection, Expressing, Plasmid Preparation, Staining, Western Blot, Control