Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Migraine‐Associated Mutation in the Na,K‐ATPase Leads to Disturbances in Cardiac Metabolism and Reduced Cardiac Function

doi: 10.1161/JAHA.121.021814

Figure Lengend Snippet: The hearts from 3‐ and 8‐month‐old α 2 +/G301R mice showed reduced expression of the Na,K‐ATPase α 2 isoform ( A , n=7; D , n=6), increased expression of Na,K‐ATPase α 1 isoform ( B , n=7; E , n=5 to 6), and no change in the expression of Na,Ca‐exchager‐1 (NCX1; C , n=7; F , n=12) in comparison with wild type (WT). The expression profile was the same in hearts of 3‐month‐old ( A – C ) and 8‐month‐old ( D – F ) mice. Upper part of each panel shows representative Western blot bands for the averaged data shown below. The representative bands are shown from the same membrane, as well as the molecular marker shown to the right. The images cropped to include molecular weights from 250 to 50 kDa, as indicated. Total protein load was detected with the stain‐free gel, and the corresponding representative bands were cropped to the same molecular weight range. Protein expression was compared using unpaired t ‐test. * P <0.05 and ** P <0.01.

Article Snippet: Antibody against the Na,Ca‐exchanger‐1 (1:200; No. ANX‐011; Alomone Labs, Israel).

Techniques: Expressing, Comparison, Western Blot, Membrane, Marker, Staining, Molecular Weight

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: 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