Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: List of primary antibodies

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques:

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: Generation of conditional renal tubular ASIC1a-knockout mice (ASIC1a fl/fl /CDH16 cre ). A Schematic diagram of constructing ASIC1a fl/fl /CDH16 cre mice; P1 and P2 were used for the PCR genotyping of exon 3 in the ASIC1a gene. B PCR genotyping of the floxed allele: primers P1 and P2 amplify a PCR product of 442 bp for the floxed allele and 281 bp for the wild-type allele, respectively. C PCR genotyping of CDH16 cre : primers amplify a PCR product of 420 bp. D Representative images of co-immunostaining ASIC1a and AQP1. ASIC1a, acid-sensing ion channel 1a; CDH16, Cadherin 16; AQP1, aquaporin 1, proximal tubular cells marker; DAPI, 4,6-diamidino-2-phenylindole, for nuclei

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Knock-Out, Immunostaining, Marker

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: Knockout of ASIC1a in the kidney epithelium attenuates renal IRI. WT, ASIC1a fl/fl , and ASIC1a fl/fl /CDH16 cre mice were subjected to renal I/R. Twenty-four hours after reperfusion, kidney and serum samples were collected. A Typical visual field of PAS staining (left panel) and pathological score calculated from PAS staining (right panel). B Levels of serum creatinine. C , D Quantification of kidney KIM-1 and NGAL (markers of renal tubular damage) mRNA levels by qPCR. E Typical image of DHE staining (left panel) and quantification of DHE fluorescence intensity (right panel). F Typical image of TUNEL staining (left panel) and the number of TUNEL-positive cells in the photographic area (right panel); TUNEL-positive cells indicated by white arrows. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Knock-Out, Staining, Fluorescence, TUNEL Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: Knockout of ASIC1a in the kidney epithelium reduces IRI-induced NLRP3 inflammasome activation by inhibiting NF-κB. A Immunohistochemistry analysis of NLRP3 expression in kidney tissues. B Percentage of positive staining for the molecule measured relative to the photographic area. C – J Western blot and densitometry analysis of NLRP3, ASC, cleaved-caspase-1 p20, GSDMD-N, IL-1β, p-NF-κB p65, and NF-κB p65 expression in kidney tissues. K Fraction of Ly6G + CD45 + cells among total CD45 + cells. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Knock-Out, Activation Assay, Immunohistochemistry, Expressing, Staining, Western Blot

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: Inhibition of ASIC1a protects HK-2 cells from H/R injury and reduces H/R-induced NLRP3 inflammasome and NF-κB activation. PcTx-1 (25 ng mL −1 ) was administered to HK-2 cells before H/R treatment. A Typical image of apoptosis measured by TUNEL in HK-2 cells. B The number of TUNEL-positive cells in the photographic area. C The cell viability of HK-2 cells assessed by CCK-8 assay. D – J Western blot and densitometry analysis of NLRP3, ASC, cleaved-caspase-1 p20, GSDMD-N, IL-1β, p-NF-κB p65, and NF-κB p65 expression in HK-2 cells. K Typical image of the NF-κB p65 nuclear translocation in HK-2 cells; nucleus NF-κB p65 indicated by purple arrows; cytoplasm NF-κB p65 indicated by red arrows. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Inhibition, Activation Assay, TUNEL Assay, CCK-8 Assay, Western Blot, Expressing, Translocation Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: ASIC1a promotes H/R-induced NLRP3 inflammasome through the NF-κB pathway. HK-2 cells were pretreated with BAY 11-7082 before H/R treatment. A CCK-8 assay indicated that a high concentration (50 μM) of BAY 11-7082 was toxic to HK-2 cells. B Western blot showed that BAY 11-7082 at 10 μM significantly reduced expression of p-NF-κB p65. C Western blot and densitometry analysis of p-NF-κB p65 and NF-κB p65 expression in HK-2 cells. D Typical image of the NF-κB p65 nuclear translocation in HK-2 cells; nucleus NF-κB p65 indicated by purple arrows; cytoplasm NF-κB p65 indicated by red arrows. E Western blot and densitometry analysis of NLRP3, ASC, cleaved-caspase-1 p20, GSDMD-N, and IL-1β expression in HK-2 cells. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: CCK-8 Assay, Concentration Assay, Western Blot, Expressing, Translocation Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: ASIC1a promotes the acidosis-induced NLRP3 inflammasome through the NF-κB pathway. HK-2 cells were pretreated with BAY 11-7082 before acidifying the medium. A Western blot of NLRP3 expression in HK-2 cells stimulated with different pH’s. B Western blot and densitometry analysis of p-NF-κB p65 and NF-κB p65 expression in HK-2 cells. C Typical image of NF-κB p65 nuclear translocation in HK-2 cells; nucleus NF-κB p65 indicated by purple arrows; cytoplasmic NF-κB p65 indicated by red arrows. D Western blot and densitometry analysis of NLRP3, ASC, cleaved-caspase-1 p20, GSDMD-N, and IL-1β expression in HK-2 cells. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Western Blot, Expressing, Translocation Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Acid-sensing ion channel 1a exacerbates renal ischemia–reperfusion injury through the NF-κB/NLRP3 inflammasome pathway

doi: 10.1007/s00109-023-02330-7

Figure Lengend Snippet: A schematic representation of the proposed model: ASIC1a is involved in renal IRI via the NF-κB/NLRP3 inflammasome. ASIC1a is activated by ischemia-induced extracellular acidosis and subsequently causes activation of NF-κB. NF-κB upregulates the expression of NLRP3 and IL-1β precursor and promotes activation of the NLRP3 inflammasome, which results in renal IRI. Inhibition of NF-κB pathway mitigates the activating effect of ASIC1a toward the NLRP3 inflammasome. Inhibiting ASIC1a by gene deletion or chemical agent protects against renal IRI. ASIC1a mediating the NLRP3 inflammasome activation by NF-κB pathway is critical in AKI. ASIC1a may represent a potential therapeutic target for AKI

Article Snippet: ASIC1a , Alomone Labs , AGP-053.

Techniques: Activation Assay, Expressing, Inhibition