primary antibody against kim 1  (R&D Systems)

Journal: The FASEB Journal

Article Title: ZXDB Drives Macrophage Inflammatory Programming in Sepsis‐Induced Acute Kidney Injury by Recruiting EIF4A3 to Enhance ACACA Translation

doi: 10.1096/fj.202502962RR

Figure Lengend Snippet: Macrophage‐Specific Deletion of Zxdb Protects Against Sepsis‐Induced Acute Kidney Injury. (A) Generation strategy for myeloid‐specific Zxdb knockout (Mac‐Zxdb‐KO) mice. (B) H&E staining of kidney sections from Sham, WT SI‐AKI, and Mac‐Zxdb‐KO SI‐AKI mice. (C‐D) Serum creatinine (Scr) (C) and blood urea nitrogen (BUN) (D) levels in Sham, WT SI‐AKI, and Mac‐Zxdb‐KO SI‐AKI mice at 24 h post‐CLP. (E) KIM‐1 immunohistochemistry of kidney sections from Sham, WT SI‐AKI, and Mac‐Zxdb‐KO SI‐AKI mice. (F) TUNEL staining of kidney sections from Sham, WT SI‐AKI, and Mac‐Zxdb‐KO SI‐AKI mice. (G) Immunofluorescence of kidney sections stained for F480, CD86, and CD206. (H) Serum IL‐1β, IL‐10, and TNF‐α levels in Sham, WT SI‐AKI, and Mac‐Zxdb‐KO SI‐AKI mice at 24 h post‐CLP. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were blocked and incubated overnight at 4°C using primary antibody against KIM‐1 (AF1817; R&D Systems, 1:200).

Techniques: Knock-Out, Staining, Immunohistochemistry, TUNEL Assay, Immunofluorescence

Journal: Renal Failure

Article Title: Magnesium hydride protects against acetaminophen-induced acute kidney injury by inhibiting TXNIP/NLRP3/nf-κb pathway

doi: 10.1080/0886022X.2024.2330629

Figure Lengend Snippet: MgH 2 ameliorates APAP-induced renal dysfunction, histological injury and oxidative stress in mice. (A, B) The levels of SCr and BUN in serum of mice were determined by corresponding kits. (C–E) The renal histological changes in mice were observed by HE staining (C), Masson staining (D) and PAS staining (E). (F, G) The protein expressions of NGAL, KIM-1 and iNOS in kidneys of mice were detected by western blotting. (H) The intracellular ROS level in renal tissues of mice was detected using DHE. The results were expressed as mean ± SEM. Statistical comparisons were performed using a Newman–Keuls test (* p < 0.05 vs. Control group, # p < 0.05 vs. APAP group).

Article Snippet: After blocking in 5% bovine serum for 2 h, the membranes were incubated at 4 °C overnight with primary antibodies against neutrophil gelatinase-associated lipocalin (NGAL; 1:1000, Cloud-Clone, China), kidney injury molecule-1 (KIM-1; 1:1000, Cloud-Clone, China), inducible nitric oxide synthase (iNOS; 1:1000, Cell Signaling Technology, USA), tumor necrosis factor-α (TNF-α; 1:1000, Servicebio, China), interleukin-1β (IL-1β; 1:1000, Cloud-Clone, China), Bcl2-antagonist of cell death (Bad; 1:1000, Cloud-Clone, China), Bcl2 associated X protein (Bax; 1:1000, Cell Signaling Technology, USA), Caspase3 (1:1000, Proteintech, USA), cytochrome C (CytC; 1:1000, Cell Signaling Technology, USA), TXNIP (1:1000), NLRP3 (1:1000, Cell Signaling Technology, USA), NF-κB p65 (1:1000, Cell Signaling Technology, USA), p-NF-κB p65 (1:1000), β-Actin (1:1000, Cell Signaling Technology, USA), β-Tubulin (1:1000, Abways, China) and GAPDH (1:1000, Servicebio, China).

Techniques: Staining, Western Blot

Journal: Renal Failure

Article Title: Magnesium hydride protects against acetaminophen-induced acute kidney injury by inhibiting TXNIP/NLRP3/nf-κb pathway

doi: 10.1080/0886022X.2024.2330629

Figure Lengend Snippet: MgH 2 alleviates APAP-induced cytotoxicity, oxidative stress, mitochondrial dysfunction and inflammation in HK-2 cells. (A-C) HK-2 Cell viability was detected by CCK-8 assay. (A) HK-2 cells was administered with different concentrations of MgH 2 (10, 20, 50, 100, 200, 400, 600, 800 and 1000 μM); (B) HK-2 cells was administered with different concentrations of APAP (5, 10, 20, 40, 60, 80, 100 mM) with or without MgH 2 (0.4 mM); (C) HK-2 cells was administered with APAP (10 mM) with or without MgH 2 (0.2, 0.4 mM). (D) The ROS level in HK-2 cells was determined by DCFH-DA. (E) The mitochondrial function of HK-2 cells was assessed by JC-1 staining. (F, G) The protein expressions of iNOS, KIM-1 and IL-1β in HK-2 cells were detected by western blotting. The results were expressed as mean ± SEM. Statistical comparisons were performed using t -test (* p < 0.05 vs. APAP group) or Newman–Keuls test (* p < 0.05 vs. Control group, # p < 0.05 vs. APAP group).

Article Snippet: After blocking in 5% bovine serum for 2 h, the membranes were incubated at 4 °C overnight with primary antibodies against neutrophil gelatinase-associated lipocalin (NGAL; 1:1000, Cloud-Clone, China), kidney injury molecule-1 (KIM-1; 1:1000, Cloud-Clone, China), inducible nitric oxide synthase (iNOS; 1:1000, Cell Signaling Technology, USA), tumor necrosis factor-α (TNF-α; 1:1000, Servicebio, China), interleukin-1β (IL-1β; 1:1000, Cloud-Clone, China), Bcl2-antagonist of cell death (Bad; 1:1000, Cloud-Clone, China), Bcl2 associated X protein (Bax; 1:1000, Cell Signaling Technology, USA), Caspase3 (1:1000, Proteintech, USA), cytochrome C (CytC; 1:1000, Cell Signaling Technology, USA), TXNIP (1:1000), NLRP3 (1:1000, Cell Signaling Technology, USA), NF-κB p65 (1:1000, Cell Signaling Technology, USA), p-NF-κB p65 (1:1000), β-Actin (1:1000, Cell Signaling Technology, USA), β-Tubulin (1:1000, Abways, China) and GAPDH (1:1000, Servicebio, China).

Techniques: CCK-8 Assay, Staining, Western Blot