Journal: International Journal of Molecular Sciences

Article Title: Kidney Injury Causes Accumulation of Renal Sodium That Modulates Renal Lymphatic Dynamics

doi: 10.3390/ijms23031428

Figure Lengend Snippet: NKCC-1 transporter expression in renal lymphatic vessels and vessels with PAN proteinuric injury. ( A ) Immunostaining of afferent renal lymphatic vessels demonstrated NKCC1 transporter expression, particularly prominent in lymphatic endothelial cells. ( B ) NKCC1 mRNA levels in extra-renal lymphatic vessels were significantly greater in PAN-injured rats vs uninjured controls. Results are mean ± SEM for 7 rats per group analyzed by unpaired t test * p < 0.05.

Article Snippet: After blocking with 2.5% normal horse serum, the tissue was incubated with anti-NKCC1 antibody (Boster Bio, 1:1000, Pleasanton, CA, USA) overnight and then incubated with anti-rabbit secondary antibody.

Techniques: Expressing, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Kidney Injury Causes Accumulation of Renal Sodium That Modulates Renal Lymphatic Dynamics

doi: 10.3390/ijms23031428

Figure Lengend Snippet: High Na + environment regulated NKCC-1 signaling pathway in lymphatic endothelial cells (LECs). ( A ) Cultured LECs exposed to a high-sodium environment showed greater expression of NKCC1 mRNA, ( B ) while expression of phosphorated NKCC-1 protein decreased. ( C ) High-sodium environment decreased protein expression of SPAK, an upstream activating kinase of NKCC1. Experiments were performed independently 3 times using 3 wells per treatment and analyzed by ANOVA followed by Dunnett multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After blocking with 2.5% normal horse serum, the tissue was incubated with anti-NKCC1 antibody (Boster Bio, 1:1000, Pleasanton, CA, USA) overnight and then incubated with anti-rabbit secondary antibody.

Techniques: Cell Culture, Expressing

Journal: Frontiers in Immunology

Article Title: Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways

doi: 10.3389/fimmu.2018.02049

Figure Lengend Snippet: Expressions of p-NKCC1 in rat lung tissues and MLE-12 cells. (A) Representative images of p-NKCC1 immunofluorescence staining (FITC-labeled green; original magnification ×400) of rat lung. Nuclei were counterstained with DAPI (blue). (B) T-NKCC1 and p-NKCC1 expressions in MLE-12 cells determined by western blot analysis ( n = 5 per group). (C) Histogram of p-NKCC1 determined by flow cytometry in MLE-12 cells ( n = 3 per group). BMT attenuated the activation of epithelial p-NKCC1 in IR-ALI. BMT, bumetanide 20-μM in HR model and 70 μg/kg in IR model; CTRL, control. Data are expressed as the means ± SD. * P < 0.05 compared with the control group; # P < 0.05 compared with the HR group.

Article Snippet: The anti-phosphorylated NKCC1 and anti-NKCC1 antibodies were custom made by GeneTex.

Techniques: Immunofluorescence, Staining, Labeling, Western Blot, Flow Cytometry, Activation Assay, Control

Journal: Frontiers in Immunology

Article Title: Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways

doi: 10.3389/fimmu.2018.02049

Figure Lengend Snippet: The mechanisms of NKCC1 modulating alveolar fluid clearance and inflammation in IR-ALI. IR stress causes phosphorylation of NKCC1 and activation of TRAF6, which result in cell swelling and inflammation of alveolar epithelium. Inhibition of NKCC1 by bumetanide reciprocally modulates epithelial TRAF6 expression. This interaction suppresses downstream p38 MAPK and NF-κB pathways, which attenuates the reduction of AFC via upregulating α-ENaC expression and reduces the alveolar inflammation.

Article Snippet: The anti-phosphorylated NKCC1 and anti-NKCC1 antibodies were custom made by GeneTex.

Techniques: Phospho-proteomics, Activation Assay, Inhibition, Expressing