Journal: International Journal of Molecular Sciences

Article Title: Improved Generation of Human Induced Pluripotent Stem Cell-Derived Cardiac Pacemaker Cells Using Novel Differentiation Protocols

doi: 10.3390/ijms23137318

Figure Lengend Snippet: Gene expression of ion channels, transporters, connexins, adrenergic and cholinergic receptors in cardiac pacemaker cells. Abundance of mRNA transcripts (in%) relative to gene expression of the housekeeping genes GAPDH/HPRT1/ACTB . ( A – C ): HCN channels. ( D ): Na-Ca exchanger NCX1 . ( E – G ): calcium channels. ( H ): sodium channel SCN5A . ( I – K ): potassium channels. ( L–N ): connexins. ( O – R ): adrenergic and cholinergic receptors. Data are provided as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, Diff. prot. A–E versus hRA. # p < 0.05, ## p < 0.01, ### p < 0.001, Diff. prot. B–E versus Diff. prot. A. Abbreviations: hRA: human right atrium, hSAN: human sinoatrial node, Diff. prot. = differentiation protocol.

Article Snippet: Subsequent primary antibodies were used: anti-HCN1 (IgG, rabbit, Alomone Labs, Ltd., Jerusalem, Israel), anti-HCN2 (IgG, rabbit, Alomone Labs, Ltd.), anti-HCN4 (IgG, rabbit, Alomone Labs, Ltd.), anti-NCX1 (IgG, rabbit, Alomone Labs, Ltd.), anti-Ca v 1.2 (IgG, mouse, Abcam, Cambridge, UK), anti-Na v 1.5 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx40 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx43 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx45 (IgG, mouse, Abcam) and anti- Tbx3 (IgG, mouse, Abcam), anti- Tbx18 (IgG, mouse, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Shox2 (IgG, mouse, Abcam) and anti-Nkx2.5 (IgG, rabbit, Abcam).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Improved Generation of Human Induced Pluripotent Stem Cell-Derived Cardiac Pacemaker Cells Using Novel Differentiation Protocols

doi: 10.3390/ijms23137318

Figure Lengend Snippet: Assessment of pacemaker-specific gene expression in cardiac pacemaker cells.

Article Snippet: Subsequent primary antibodies were used: anti-HCN1 (IgG, rabbit, Alomone Labs, Ltd., Jerusalem, Israel), anti-HCN2 (IgG, rabbit, Alomone Labs, Ltd.), anti-HCN4 (IgG, rabbit, Alomone Labs, Ltd.), anti-NCX1 (IgG, rabbit, Alomone Labs, Ltd.), anti-Ca v 1.2 (IgG, mouse, Abcam, Cambridge, UK), anti-Na v 1.5 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx40 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx43 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx45 (IgG, mouse, Abcam) and anti- Tbx3 (IgG, mouse, Abcam), anti- Tbx18 (IgG, mouse, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Shox2 (IgG, mouse, Abcam) and anti-Nkx2.5 (IgG, rabbit, Abcam).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Improved Generation of Human Induced Pluripotent Stem Cell-Derived Cardiac Pacemaker Cells Using Novel Differentiation Protocols

doi: 10.3390/ijms23137318

Figure Lengend Snippet: Overview of the used TaqMan probes and primers.

Article Snippet: Subsequent primary antibodies were used: anti-HCN1 (IgG, rabbit, Alomone Labs, Ltd., Jerusalem, Israel), anti-HCN2 (IgG, rabbit, Alomone Labs, Ltd.), anti-HCN4 (IgG, rabbit, Alomone Labs, Ltd.), anti-NCX1 (IgG, rabbit, Alomone Labs, Ltd.), anti-Ca v 1.2 (IgG, mouse, Abcam, Cambridge, UK), anti-Na v 1.5 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx40 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx43 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx45 (IgG, mouse, Abcam) and anti- Tbx3 (IgG, mouse, Abcam), anti- Tbx18 (IgG, mouse, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Shox2 (IgG, mouse, Abcam) and anti-Nkx2.5 (IgG, rabbit, Abcam).

Techniques:

Journal: Neurobiology of disease

Article Title: Prolonged NCX activation prevents SOD1 accumulation, reduces neuroinflammation, ameliorates motor behavior and prolongs survival in a ALS mouse model.

doi: 10.1016/j.nbd.2021.105480

Figure Lengend Snippet: Fig. 1. Evaluation of NCX1and NCX2 transcripts and protein expressions in spinal cord tissues from SOD1G93Amice compared to age-matched healthy controls (Wt). (A) and (B) show mRNA levels and protein of NCX1 at 2 months and 4.5 months of age in spinal cord of Wt and SOD1G93Amice. mRNA expression levels were normalized for beta-glucuronidase (GUSB). Protein expression levels were normalized with β-actin. (C) and (D) show mRNA and proteins levels of NCX2 at 2 months and 4.5 months of age in spinal cord of Wt and SOD1G93Amice. Real time PCR, data are expressed as mean ± SEM (n = 3/6 for each group). Western blotting analysis, data are expressed as mean ± SEM (n = 3/5 for each group). Statistically significant differences among means were determined by one-way ANOVA followed by Newman Keuls’s correction for multiple comparisons test: * p < 0.05, SOD1G93A vs. Wt; ^ p < 0.05, Wt 4.5 months vs. Wt 2 months.

Article Snippet: Rabbit polyclonal antibody against NCX1 (1:1000 dilution, Swant, Bellinzona, Switzerland), rabbit polyclonal anti-NCX2 (1:500 dilution, ALPHA-DIAGNOSTIC int.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Neurobiology of disease

Article Title: Prolonged NCX activation prevents SOD1 accumulation, reduces neuroinflammation, ameliorates motor behavior and prolongs survival in a ALS mouse model.

doi: 10.1016/j.nbd.2021.105480

Figure Lengend Snippet: Fig. 2. NCX1 and 2 immunolocalization, expression and quantification of NCX activity in SOD1G93A mice treated to neurounina. (A) Quantification of NCX activity as [Ca2+]i increase induced by Na + −free perfusion in Fura-2 AM-loaded spinal cord synaptosomes of adult wild-type (Wt) mice treated with vehicle, Wt mice treated with neurounina and symptomatic SOD1G93A mice treated with vehicle or neurounina. P values were obtained using one-way ANOVA with Newman Keuls’s correction for multiple comparisons. *P < 0.05 vs. wild-type vehicle and wild-type neurounina; ^P < 0.05 vs. SOD1G93A mice vehicle; (B) Representative western blotting and quantification of the effect of neurounina on NCX1–2 protein expression, arbitrary units (AU), in spinal cord. Statistically significant differences among means were determined by one-way ANOVA followed by Newman Keuls’s correction for multiple comparisons. The β-actin expression level was used for normalization. Data are expressed as mean ± SEM (n = 4/5 for each group). * p < 0.05, Wt vs SOD1G93A mice. Double labeling of NCX1 and NCX2 with SMI-32 in spinal cord of Wt mice + vehicle (C a-d, D a-d), Wt mice + neurounina (C e-h, D e-h) SOD1G93A mice + vehicle (C i-l, D i-l) SOD1G93A mice + neurounina (C m-p, Dm-p) Scale bar 20μm.

Article Snippet: Rabbit polyclonal antibody against NCX1 (1:1000 dilution, Swant, Bellinzona, Switzerland), rabbit polyclonal anti-NCX2 (1:500 dilution, ALPHA-DIAGNOSTIC int.

Techniques: Expressing, Activity Assay, Western Blot, Labeling

Journal: Journal of Molecular Histology

Article Title: Neuroprotection in neonatal Hypoxia-ischaemia: melatonin targets NCX1 to inhibit mitochondrial autophagy via the PINK1-Parkin pathway

doi: 10.1007/s10735-025-10601-5

Figure Lengend Snippet: Immunohistochemical and simple Western blot analysis to evaluate the effect of melatonin on NCX1 expression activity. a Immunohistochemistry was used to assess the expression levels of NCX1 in neonatal rats at 7, 14, and 21 days. In the Sham group, normal NCX1 protein expression was observed in the brain. Compared with the Sham group, the HI group showed significantly greater NCX1-positive staining at 7 days of age after the brain injury. However, the positive results for NCX1 were diminished at 14 and 21 days of age. In contrast, strong positive expression was demonstrated in the Mel group at 14 and 21 days of age. Compared with the Mel group, the SN6 group exhibited high expression at 7 days of age and low expression at 14 and 21 days of age; scale bar = 20 µm, b Average optical density (AOD) of NCX1 staining in each group (n = 6), c – d NCX1 protein expression exhibited dynamic changes in the HI group, with an increase observed on postnatal Day 7 compared with that in the Sham group but a significant decrease on Days 14 and 21. This trend was reversed in the Mel group, whose expression was lower than that in the HI group on Day 7 but significantly greater on Days 14 and 21. In the SN6 group, expression was upregulated on Day 7 but decreased at subsequent time points (n = 4), e There were no significant differences in the expression of NCX1 or autophagy-related proteins among the S, S + M, and S + M + N groups (n = 3); the data represent the mean ± SEM; ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The primary antibodies used included: Rabbit anti-NCX1 polyclonal antibody (1:20, 0.2 μg/μL, #28447-1-AP; Proteintech, Wuhan, China) Rabbit anti-PLP monoclonal antibody (1:10, 2 μg/μL, EPR23504-106, #ab254363; Abcam, Cambridge, UK) Rabbit anti-LC3B monoclonal antibody (1:100, 0.2 μg/μL, EPR18709 , #ab192890; Abcam) Mouse anti-MAG monoclonal antibody (1:20, 2 μg/μL, sc-166849; Santa Cruz Biotechnology, Dallas, TX, USA) Mouse anti-PINK1 monoclonal antibody (1:100, 2 μg/μL, sc-517353; Santa Cruz Biotechnology) Mouse anti-Parkin monoclonal antibody (1:20, 2 μg/μL, sc-32282; Santa Cruz Biotechnology)

Techniques: Immunohistochemical staining, Simple Western, Expressing, Activity Assay, Immunohistochemistry, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: Visinin-like protein 1 disrupts calcium homeostasis and promotes atrial fibrillation in human and rodent models

doi: 10.1038/s41392-026-02615-6

Figure Lengend Snippet: Sodium-calcium exchanger (NCX-1) is identified as a key downstream target for VILIP-1. a Schematic workflow illustrating immunoprecipitation-mass spectrometry (left panel) and proximity labeling experiments using VILIP-1-V5-miniTurbo under varying Ca 2+ concentrations (right panel) to identify VILIP-1 interactors. b MaSigPro (R package) identified four dynamic VILIP-1-proximity labeling patterns in response to increased Ca 2+ concentrations. Proteins in Cluster 1 were identified as VILIP-1’s candidate interactors based on their progressive enrichment in VILIP-1-V5-miniTurbo-mediated biotinylation profiles with increasing Ca 2+ concentrations. c Venn diagram illustrating the overlap between interactors identified by co-immunoprecipitation and proximity labeling, confirming high-confidence candidates. d Western blot analysis of biotinylated proteins from VILIP-1-V5-miniTurbo proximity labeling, showing Ca 2+ -dependent enrichment of NCX-1. e Confocal microscopy images confirming the localization of biotinylated VILIP-1-interacting proteins (within a 10 nm radius), NCX-1, in neonatal rat cardiomyocytes (NRCMs) expressing VILIP-1-V5-miniTurbo. Cells were treated with 500 μM biotin for 13 min at 37 °C, and biotinylated proteins were visualized using Neutravidin–Alexa Fluor 488 staining. f Representative immunoblots showing co-immunoprecipitation of VILIP-1 and NCX-1 in rat atrial lysate, demonstrating their interaction. g Western blot analysis of in vitro GST pull-down experiments from VILIP-1-GST and HA-NCX-1, confirming the direct interaction between VILIP-1 and NCX-1. h Confocal microscopy images displaying colocalization of VILIP-1 and NCX-1 in rat atrial cardiomyocytes

Article Snippet: After blocking with QuickBlock Blocking Buffer (Beyotime Biotechnology, China), the membranes were incubated overnight at 4 °C with primary antibodies, including NCX-1 (Proteintech, USA), VILIP-1 (Proteintech, USA), β-ACTIN (Santa Cruz, USA) and GAPDH (Proteintech, USA), all diluted in the same blocking buffer.

Techniques: Immunoprecipitation, Mass Spectrometry, Labeling, Western Blot, Confocal Microscopy, Expressing, Staining, In Vitro

Journal: Signal Transduction and Targeted Therapy

Article Title: Visinin-like protein 1 disrupts calcium homeostasis and promotes atrial fibrillation in human and rodent models

doi: 10.1038/s41392-026-02615-6

Figure Lengend Snippet: Atrial cardiomyocyte-specific overexpression of VILIP-1 increases atrial fibrillation (AF) susceptibility through enhancing the membrane expression and function of NCX-1. a Western blot analysis of VILIP-1 and NCX-1 protein levels in the atria of VILIP-1-OE and Ctrl mice. Left: representative immunoblot images. Right: quantification of protein levels normalized to GAPDH. * P < 0.05, Student’s t test, n = 3 per group. b Capillary western immunoassay results showed elevated NCX-1 and VILIP-1 protein levels in membrane fractions of atria from Ctrl and VILIP-1-OE mice. Quantification was performed using the Total Protein Detection Module (DM-TP01, ProteinSimple, USA). * P < 0.05, Student’s t test, n = 4 per group. c Whole-cell patch-clamp recordings from isolated atrial cardiomyocytes illustrating a significant increase in the I NCX current density in VILIP-1-OE mice compared to Ctrl. * P < 0.05, Student’s t test, n = 14 per group. d Confocal Ca 2+ imaging of atrial cardiomyocytes from VILIP-1-OE mice pretreated with vehicle or SEA0400. Left: representative Ca 2+ wave recordings. Right: quantification of the frequency of spontaneous Ca 2+ waves, showing reduced spontaneous Ca 2+ waves upon SEA0400 treatment. * P < 0.05, Mann-Whitney test, n = 13-18 per group. e Electrophysiological recordings of surface ECG and intracardiac electrograms in VILIP-1-OE mice. Left: representative traces of pacing-induced AF episodes in VILIP-1-OE mice with or without NCX-1 inhibition using SEA0400. Right: quantification of AF incidence and duration, showing a significant reduction in AF susceptibility upon SEA0400 treatment. * P < 0.05, Mann-Whitney test or Fisher’s exact test, n = 7–8 per group

Article Snippet: After blocking with QuickBlock Blocking Buffer (Beyotime Biotechnology, China), the membranes were incubated overnight at 4 °C with primary antibodies, including NCX-1 (Proteintech, USA), VILIP-1 (Proteintech, USA), β-ACTIN (Santa Cruz, USA) and GAPDH (Proteintech, USA), all diluted in the same blocking buffer.

Techniques: Over Expression, Membrane, Expressing, Western Blot, Patch Clamp, Isolation, Imaging, MANN-WHITNEY, Inhibition

Journal: Signal Transduction and Targeted Therapy

Article Title: Visinin-like protein 1 disrupts calcium homeostasis and promotes atrial fibrillation in human and rodent models

doi: 10.1038/s41392-026-02615-6

Figure Lengend Snippet: Inhibition of VILIP-1 myristoylation rescues atrial fibrillation (AF) susceptibility in rat AF models. a Capillary western immunoassay analysis and quantification of NCX-1 expression in atrial membrane fractions from AF rat models treated with vehicle or Desloratadine. Quantification was performed using the Total Protein Detection Module (DM-TP01, ProteinSimple, USA). * P < 0.05, Student’s t test, n = 4 per group. b Whole-cell patch-clamp recordings demonstrate that AF rats treated with vehicle exhibit significantly increased I NCX current density compared to the control, while desloratadine treatment restores the I NCX current density to control levels. * P < 0.05, one-way ANOVA, n = 14–18 per group. c Confocal Ca 2+ imaging of atrial cardiomyocytes from rat AF models pretreated with vehicle or desloratadine. Left: representative Ca 2+ wave recordings. Right: quantification of the frequency of spontaneous Ca 2+ waves, showing reduced Ca 2+ waves upon desloratadine treatment. * P < 0.05, Mann-Whitney test, n = 17 per group. d Electrophysiological recordings of surface ECG and intracardiac electrograms in pacing-induced AF rats. Left: representative traces showing AF episodes in vehicle-pretreated versus 5 mg/kg desloratadine-pretreated rats. Right: quantification of AF incidence and duration, demonstrating a significant reduction in AF susceptibility following desloratadine administration. * P < 0.05, Mann-Whitney test or Fisher’s exact test, n = 6–8 per group

Article Snippet: After blocking with QuickBlock Blocking Buffer (Beyotime Biotechnology, China), the membranes were incubated overnight at 4 °C with primary antibodies, including NCX-1 (Proteintech, USA), VILIP-1 (Proteintech, USA), β-ACTIN (Santa Cruz, USA) and GAPDH (Proteintech, USA), all diluted in the same blocking buffer.

Techniques: Inhibition, Western Blot, Expressing, Membrane, Patch Clamp, Control, Imaging, MANN-WHITNEY

Journal: Signal Transduction and Targeted Therapy

Article Title: Visinin-like protein 1 disrupts calcium homeostasis and promotes atrial fibrillation in human and rodent models

doi: 10.1038/s41392-026-02615-6

Figure Lengend Snippet: VILIP-1 inhibition alleviates Ca 2+ leakage and prevents atrial fibrillation (AF) inducibility in both human patients and rat AF models. a Molecular docking simulation demonstrating the interaction between repaglinide and VILIP-1 (docking score = -318.2), supporting its potential as a VILIP-1 inhibitor. b Thermal shift assay (TSA) analysis showing VILIP-1 protein stabilization following 10 μM repaglinide treatment at varying temperatures. c Binding affinities of repaglinide to VILIP-1 measured by surface plasmon resonance (SPR) assays. d Capillary western immunoassay demonstrating a reduction in NCX-1 expression in membrane fractions of atria from pacing-induced AF rat models treated with repaglinide. Quantification was performed using the Total Protein Detection Module (DM-TP01, ProteinSimple, USA). * P < 0.05, Mann-Whitney test, n = 4 per group. e Immunofluorescence staining of VILIP-1 and NCX-1 in atria from AF rat models pretreated with vehicle or repaglinide, showing reduced NCX-1 surface expression upon repaglinide treatment. f Confocal Ca 2+ imaging of atrial cardiomyocytes from AF rat models pretreated with vehicle or repaglinide. Top: representative Ca 2+ wave recordings. Bottom: quantification of the frequency of spontaneous Ca 2+ waves, demonstrating a significant reduction in spontaneous Ca 2+ waves following repaglinide treatment. * P < 0.05, Mann-Whitney test, n = 15–16 per group. g Electrophysiological recordings of surface ECG and intracardiac electrograms in pacing-induced AF rats pretreated with vehicle or repaglinide. Top: representative traces showing a decrease in pacing-induced AF episodes following repaglinide administration. Bottom: quantification of AF incidence and duration. * P < 0.05, Mann-Whitney test or Fisher’s exact test, n = 6 – 7 per group. h Capillary western immunoassay analysis of NCX-1 expression in membrane fractions of human atria, showing decreased NCX-1 surface expression following repaglinide treatment. Quantification was performed using the Total Protein Detection Module (DM-TP01, ProteinSimple, USA). * P < 0.05, Mann-Whitney test, n = 6-8 per group. i Confocal Ca 2+ imaging of atrial cardiomyocytes from human AF patients pretreated with vehicle or 10 μM repaglinide. Left: representative Ca 2+ wave recordings. Right: quantification of the frequency of spontaneous Ca 2+ waves, demonstrating a significant reduction in spontaneous Ca 2+ waves following repaglinide treatment. * P < 0.05, Mann-Whitney test, n = 17–19 per group

Article Snippet: After blocking with QuickBlock Blocking Buffer (Beyotime Biotechnology, China), the membranes were incubated overnight at 4 °C with primary antibodies, including NCX-1 (Proteintech, USA), VILIP-1 (Proteintech, USA), β-ACTIN (Santa Cruz, USA) and GAPDH (Proteintech, USA), all diluted in the same blocking buffer.

Techniques: Inhibition, Thermal Shift Assay, Binding Assay, SPR Assay, Western Blot, Expressing, Membrane, MANN-WHITNEY, Immunofluorescence, Staining, Imaging

Journal: Signal Transduction and Targeted Therapy

Article Title: Visinin-like protein 1 disrupts calcium homeostasis and promotes atrial fibrillation in human and rodent models

doi: 10.1038/s41392-026-02615-6

Figure Lengend Snippet: Upregulated VILIP-1 promotes atrial fibrillation (AF) through a myristoylation-dependent pathway. In pathological conditions, upregulated VILIP-1 binds Ca 2+ , triggering myristoylation-dependent membrane anchoring. At the membrane, VILIP-1 directly increases the surface membrane expression of NCX-1 and enhances its function. The sustained Ca 2+ overload combined with intracellular Na + accumulation drives NCX-1 reverse-mode activation, exacerbating both Ca 2+ influx and cytoplasmic Ca 2+ accumulation. The resulting Ca 2+ overload reinforces VILIP-1 membrane targeting by promoting its myristoylation, creating a self-perpetuating cycle driving NCX-1 hyperactivity and AF progression. The schematic illustration was created in Adobe Illustrator 2024

Article Snippet: After blocking with QuickBlock Blocking Buffer (Beyotime Biotechnology, China), the membranes were incubated overnight at 4 °C with primary antibodies, including NCX-1 (Proteintech, USA), VILIP-1 (Proteintech, USA), β-ACTIN (Santa Cruz, USA) and GAPDH (Proteintech, USA), all diluted in the same blocking buffer.

Techniques: Membrane, Expressing, Activation Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Targeting CXCR2 ameliorated tacrolimus-induced nephrotoxicity by alleviating overactivation of PI3K/AKT/mTOR pathway and calcium overload.

doi: 10.1016/j.biopha.2024.117526

Figure Lengend Snippet: Fig. 3. Fibrosis and calcium overload occurred in tacrolimus-induced nephrotoxicity. (A) Masson staining. (B) Immumohistochemical staining of vimentin. (C) Proteomic analysis. (D) Immumohistochemical staining of NCX1. (E) Immunofluorescent staining of CaBP28K. (F) Western blotting of E-cadherin, α-SMA, vimentin, NCX1, and CaBP28K. (G) The expression of E-cadherin. (H) The expression of α-SMA. (I) The expression of vimentin. (J) The expression of NCX1. (K) The expression of CaBP28K. **P < 0.01 vs N. N, control group. T, tacrolimus intervention group. Scale bar, 200 µm.

Article Snippet: Immumohistochemical staining was used for analyzing the expression of vimentin, NCX1, where antibody against vimentin (GB11192) was purchased from Servicebio Biotechnology (Wuhan) Co., Ltd and antibody against NCX1 (TD9932) was purchased from Abmart Medical Technology (Shanghai) Co., Ltd. Paraffin section dewaxed to water, antigen repair, blocking endogenous peroxidase, adding corresponding antibodies, dyeing nucleus, sealing, and the positive expression of antibody was brownish-yellow.

Techniques: Staining, Western Blot, Expressing, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Targeting CXCR2 ameliorated tacrolimus-induced nephrotoxicity by alleviating overactivation of PI3K/AKT/mTOR pathway and calcium overload.

doi: 10.1016/j.biopha.2024.117526

Figure Lengend Snippet: Fig. 7. Blocking CXCR2 reversed fibrosis and calcium overload associated proteins in tacrolimus-induced nephrotoxicity. (A) Western blotting of E-cad herin, α-SMA, vimentin, NCX1, and CaBP28K. (B) The expression of E-cadherin. (C) The expression of α-SMA. (D) The expression of vimentin. (E) The expression of NCX1. (F) The expression of CaBP28K. (G) Immumohistochemical staining of NCX1 and immunofluorescent staining of CaBP28K. *P < 0.05, **P < 0.01 vs N; #P < 0.05, ##P < 0.01 vs T; N, control group. T, tacrolimus intervention group. C, CXCR2 antagonist group. Scale bar, 200 µm.

Article Snippet: Immumohistochemical staining was used for analyzing the expression of vimentin, NCX1, where antibody against vimentin (GB11192) was purchased from Servicebio Biotechnology (Wuhan) Co., Ltd and antibody against NCX1 (TD9932) was purchased from Abmart Medical Technology (Shanghai) Co., Ltd. Paraffin section dewaxed to water, antigen repair, blocking endogenous peroxidase, adding corresponding antibodies, dyeing nucleus, sealing, and the positive expression of antibody was brownish-yellow.

Techniques: Blocking Assay, Western Blot, Expressing, Staining, Control

Journal: Biological Research

Article Title: Inhibition of forward and reverse transport of Ca 2+ via Na + /Ca 2+ exchangers (NCX) prevents sperm capacitation

doi: 10.1186/s40659-024-00535-9

Figure Lengend Snippet: Identification of NCX isoforms. Identification of NCX1, NCX2 and NCX3 in porcine sperm. Representative immunoblots using sperm samples from different boars with A ) anti-NCX1, ( B ) anti-NCX2 and ( C ) anti-NCX3 antibodies, and their respective blocking peptides

Article Snippet: ; Basel, Switzerland) at room temperature and agitation for 1 h. Membranes were subsequently incubated with specific primary antibodies against NCX1 (SLC8A1), NCX2 (SLC8A2), or NCX3 (SLC8A3) (Alomone Labs, Jerusalem, Israel), which were previously diluted in blocking solution at 1:2,000 (v: v), at 4 °C overnight under agitation.

Techniques: Western Blot, Blocking Assay

Journal: Biological Research

Article Title: Inhibition of forward and reverse transport of Ca 2+ via Na + /Ca 2+ exchangers (NCX) prevents sperm capacitation

doi: 10.1186/s40659-024-00535-9

Figure Lengend Snippet: Immunolocalization of NCX1 isoform. Localization of NCX1 in the plasma membrane of porcine sperm ( A – C ), and after the peptide competition assay ( D – F ). NCX1 appears stained in green (Alexa Fluor 488) and nuclei in blue (DAPI; 4′6′-diamidion-2-phenylindole). Scale bar: 15 μm

Article Snippet: ; Basel, Switzerland) at room temperature and agitation for 1 h. Membranes were subsequently incubated with specific primary antibodies against NCX1 (SLC8A1), NCX2 (SLC8A2), or NCX3 (SLC8A3) (Alomone Labs, Jerusalem, Israel), which were previously diluted in blocking solution at 1:2,000 (v: v), at 4 °C overnight under agitation.

Techniques: Clinical Proteomics, Membrane, Competitive Binding Assay, Staining