renal tubular epithelial cell line hk 2  (ATCC)

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The effects of NTN1 on the apoptosis, inflammatory levels and ROS generation in LPS-treated HK-2 cells (A–B) after the transfection of NTN1 overexpression plasmid or si-NTN1, the expression of NTN1 in the blank, OE-NC, OE-NTN1, si-NC, and si-NTN1 groups was determined by qRT-PCR. GAPDH served as the internal control. (C–D) the apoptosis rate of HK-2 cells or RPTEC/TERT1 in the OE-NC, OE-NTN1, si-NC, and si-NTN1 groups was assessed by flow cytometry. (E–G) the levels of TNF-α, IL-6 and IL-1β in the OE-NC, OE-NTN1, si-NC, and si-NTN1 groups were determined by ELISA. (H) The level of ROS generation in each group was detected by DCFH-DA reagent. The data are presented as the mean±standard deviation of three independent experiments; *** p<0.001 vs. OE-NC; ### p<0.001 vs. si-NC. Abbreviation: NTN1, netrin 1; ROS, reactive oxygen species; LPS, lipopolysaccharides; OE-NTN1, NTN1 overexpression; si-NTN1, silenced NTN1; NC, negative control; qRT-PCR, quantitative real-time PCR; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; ELISA, enzyme-linked immunosorbent assay; DCFH-DA, 2,7-Dichlorodi-hydrofluorescein diacetate.

Article Snippet: Renal tubular epithelial cell line HK-2 (AW-CELLS-H0142, Anweisci, China), and RPTEC/TERT1 cells (CRL-4031, ATCC, Virginia, MA, USA) were placed in Minimum Essential Medium (MEM, BC-M-019, Sbjbio, China) enriched with 10 % fetal bovine serum (FBS, FCS500, ExCell, China) at 37 °C with 5 % CO 2 .

Techniques: Transfection, Over Expression, Plasmid Preparation, Expressing, Quantitative RT-PCR, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Standard Deviation, Negative Control, Real-time Polymerase Chain Reaction

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The effects of MAP1B phosphorylation inhibitor on the apoptosis and autophagy in LPS-treated HK-2 cells (A) after the treatment of MAP1B phosphorylation inhibitor, the apoptosis of HK-2 cells in the OE-NC, OE-NTN1 and OE-NTN1+inhibitor groups was evaluated by flow cytometry. (B) The protein expression of Atg5 and the ratio of LC3II/LC3I were determined by western blot. GAPDH served as the internal control. The data are presented as the mean ± standard deviation of three independent experiments; *** p<0.001 vs. OE-NC; &&& p<0.001 vs. OE-NTN1. Abbreviation: MAP1B, microtubule associated protein 1B; LPS, lipopolysaccharides; NTN1, netrin 1; OE-NTN1, NTN1 overexpression; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Renal tubular epithelial cell line HK-2 (AW-CELLS-H0142, Anweisci, China), and RPTEC/TERT1 cells (CRL-4031, ATCC, Virginia, MA, USA) were placed in Minimum Essential Medium (MEM, BC-M-019, Sbjbio, China) enriched with 10 % fetal bovine serum (FBS, FCS500, ExCell, China) at 37 °C with 5 % CO 2 .

Techniques: Phospho-proteomics, Flow Cytometry, Expressing, Western Blot, Control, Standard Deviation, Over Expression, Negative Control

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The effects of MAP1B and DAPK1 on the apoptosis and ROS generation of LPS-treated HK-2 cells (A) Co-IP assay was performed to detect the relationship between MAP1B and DAPK1. (B–C) after the transfection of si-DAPK1 (B) or OE-MAP1B (C), the mRNA expression of DAPK1 or MAP1B in HK-2 cells was tested by qRT-PCR. GAPDH served as the internal control. (D) flow cytometry was used to determine the apoptosis rate of LPS-treated HK-2 cells in the si-NC+OE-NC, si-DAPK1+OE-NC, si-NC+OE-MAP1B and si-DAPK1+OE-MAP1B groups. (E) the ROS generation in each group was detected by DCFH-DA reagent. The data are presented as the mean±standard deviation of three independent experiments; ### p<0.001 vs. si-NC; +++ p<0.001 vs. OE-NC; *** p<0.001 vs. si-NC+OE-NC; ˆˆˆ p<0.001 vs. si-DAPK1+OE-NC; &&& p<0.001 vs. si-NC+OE-MAP1B. Abbreviation: MAP1B, microtubule associated protein 1B; DAPK1, death associated protein kinase 1; ROS, reactive oxygen species; LPS, lipopolysaccharides; si-DAPK1, silenced DAPK1; NC, negative control; qRT-PCR, quantitative real-time PCR; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DCFH-DA, 2,7-Dichlorodi-hydrofluorescein diacetate.

Article Snippet: Renal tubular epithelial cell line HK-2 (AW-CELLS-H0142, Anweisci, China), and RPTEC/TERT1 cells (CRL-4031, ATCC, Virginia, MA, USA) were placed in Minimum Essential Medium (MEM, BC-M-019, Sbjbio, China) enriched with 10 % fetal bovine serum (FBS, FCS500, ExCell, China) at 37 °C with 5 % CO 2 .

Techniques: Co-Immunoprecipitation Assay, Transfection, Expressing, Quantitative RT-PCR, Control, Flow Cytometry, Standard Deviation, Negative Control, Real-time Polymerase Chain Reaction

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The effects of MAP1B and DAPK1 on the membrane blebbing and autophagy of LPS-treated HK-2 cells (A) the membrane blebbing of LPS-treated HK-2 cells in the si-NC+OE-NC, si-DAPK1+OE-NC, si-NC+OE-MAP1B and si-DAPK1+OE-MAP1B groups was observed under the microscope (magnification × 400, scale bar=100 μm). (B) The protein expression of Atg5 and the ratio of LC3II/LC3I in each group were determined by western blot. GAPDH served as the internal control. The data are presented as the mean±standard deviation of three independent experiments; ** p<0.01, *** p<0.001 vs. si-NC+OE-NC; ˆˆ p<0.01, ˆˆˆ p<0.001 vs. si-DAPK1+OE-NC; &&& p<0.001 vs. si-NC+OE-MAP1B; ++ p<0.01, +++ p<0.001 vs si-NC+OE-NC+BafA1; ΔΔ p <0.01, ΔΔΔ p <0.001 vs. si-DAPK1+OE-NC+BafA1; ### p<0.001 vs. si-NC+OE-MAP1B+BafA1; $$$ p<0.001vs. si-DAPK1+OE-MAP1B. Abbreviation: MAP1B, microtubule associated protein 1B; DAPK1, death associated protein kinase 1; LPS, lipopolysaccharides; si-DAPK1, silenced DAPK1; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Renal tubular epithelial cell line HK-2 (AW-CELLS-H0142, Anweisci, China), and RPTEC/TERT1 cells (CRL-4031, ATCC, Virginia, MA, USA) were placed in Minimum Essential Medium (MEM, BC-M-019, Sbjbio, China) enriched with 10 % fetal bovine serum (FBS, FCS500, ExCell, China) at 37 °C with 5 % CO 2 .

Techniques: Membrane, Microscopy, Expressing, Western Blot, Control, Standard Deviation, Negative Control

Journal: Materials Today Bio

Article Title: Ferrostatin-1-loaded black phosphorus quantum dots (BPQD@Fer-1) nanodelivery system attenuates T cell-mediated rejection after kidney transplantation

doi: 10.1016/j.mtbio.2026.103292

Figure Lengend Snippet: Schematic illustration of the construction of BPQD@Fer-1 nanoparticles and their application in alleviating TCMR in kidney transplantation. Top panel: Synthesis of black phosphorus quantum dots loaded with Ferrostatin-1 (BPQD@Fer-1) via liquid-phase exfoliation of bulk BP in NMP. The nanoparticles exhibit intrinsic ROS-scavenging capabilities by neutralizing free radicals (e.g., ⋅O 2 −and ⋅OH) through electron (e − ) transfer. Middle panel: In vivo application in a murine kidney transplantation model. Donor kidneys are subjected to cold ischemia and subsequently transplanted. Intravenously administered BPQD@Fer-1 selectively accumulate in the tubular epithelial cells of the kidney allograft. Bottom panel: Intracellular mechanisms and immune microenvironment remodeling. (Left, TCMR group): Severe oxidative stress upregulates intracellular ROS and lipid peroxidation (LPO), triggering ferroptosis in tubular epithelial cells. This leads to the massive release of damage-associated molecular patterns (DAMPs), including high mobility group box 1 (HMGB1), calreticulin (CRT), lactate dehydrogenase (LDH), and adenosine triphosphate (ATP), which subsequently recruit and activate CD8 + T cells, resulting in the upregulation of cytotoxic and pro-inflammatory cytokines (GzmB, IL-2, TNF-α, and IFN-γ). (Right, BPQD@Fer-1 group): The nanoparticles efficiently scavenge ROS, suppress LPO, and block the ferroptotic cascade. The consequent inhibition of DAMPs release restricts CD8 + T cell-mediated cytotoxicity and downregulates the inflammatory cytokine storm, ultimately preserving the kidney allograft.

Article Snippet: Human renal tubular epithelial cell line (HK-2) and rat renal tubular epithelial cell line (NRK52E) were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Transplantation Assay, In Vivo, Blocking Assay, Inhibition, Preserving

Journal: Materials Today Bio

Article Title: Ferrostatin-1-loaded black phosphorus quantum dots (BPQD@Fer-1) nanodelivery system attenuates T cell-mediated rejection after kidney transplantation

doi: 10.1016/j.mtbio.2026.103292

Figure Lengend Snippet: Single-cell landscape reveals ferroptosis-associated epithelial states and enhanced T-cell activation in TCMR. (A) UMAP visualization of all single cells colored by sample origin (left panel) and cell cluster (right panel). (B) Dot plot showing the expression of canonical marker genes used for cell type annotation across major renal epithelial, immune, and stromal populations. Dot size represents the percentage of cells expressing each gene, and color intensity indicates scaled average expression. (C) UMAP plot annotated by cell type. (D) UMAP plots split by experimental condition (CTRL and TCMR), illustrating comparable global cellular architecture across conditions. (E) Boxplot showing ferroptosis module scores in epithelial cells from CTRL and TCMR groups. Each dot represents a single cell. (F) Ferroptosis scores across epithelial subtypes, including proximal tubule, thick ascending limb, collecting duct principal and intercalated cells, thin limb, and PEC subsets, stratified by condition (CTRL vs TCMR). (G) Heatmap showing average expression of ferroptosis-related genes across epithelial cell types and conditions. Expression values are scaled by gene to highlight relative differences. (H) Boxplot of DAMPs module scores in epithelial cells stratified by ferroptosis state (Ferro_High vs Ferro_Low). (I) Dot plot visualizing the expression profiles of DAMPs-associated genes in epithelial cells, stratified by their ferroptosis states. Dot diameter corresponds to the proportion of cells expressing the target gene, while the color gradient indicates the average expression intensity. (J) Boxplots delineating the variations in T-cell activation scores among the three defined ferroptosis subgroups (Ferro_High, Ferro_Med, and Ferro_Low). (K) Dot plot displaying the expression dynamics of T-cell-related markers across different ferroptosis cohorts. The cellular fraction expressing each gene is represented by dot size, with the mean expression level denoted by color scaling. (L) Boxplot showing cytotoxicity scores in T cells across ferroptosis groups.

Article Snippet: Human renal tubular epithelial cell line (HK-2) and rat renal tubular epithelial cell line (NRK52E) were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Single Cell, Activation Assay, Expressing, Marker

Journal: Comprehensive Physiology

Article Title: The cardiac circadian clock regulates rhythms in peripheral tissues via Fibulin 5

doi: 10.1002/cph4.70147

Figure Lengend Snippet: Treatment of synchronized kidney and muscle cells with recombinant Fbln5 (10ng/mL) disrupts rhythmic gene expression. A,B) Disrupted expression of Clock and Bmal1 but not C) Per2 in mouse myotube C2C12 cells with Fbln5 treatment. D,E) Disrupted expression of Bmal1 and Per1 but not F) Clock in human renal proximal tubule endothelial cells (RPTEC) with Fbln5. n=3 repetitions with biological triplicate. *p<0.05 Vehicle vs Fbln5. Cells were synchronized with dexamethasone prior to treatment with Fbln5 or vehicle.

Article Snippet: The human Renal Proximal Tubule Epithelial Cell (RPTEC) immortalized cell line (ATCC, PCS-400–010) was cultured in complete growth medium consisting of the base medium containing DMEM: F12 Medium (ATCC 30–2006) and the RPTEC Growth Kit components (ATCC ACS-4007) consisting of 5 pM triiodo-L-thyronine, 10 ng/mL recombinant human EGF, 3.5 μg/mL ascorbic acid, 5.0 μg/mL human transferrin, 5.0 μg/mL insulin, 25 ng/mL prostaglandin E 1 , 25 ng/mL hydrocortisone, 8.65 ng/mL sodium selenite and 1.2 mg/mL sodium bicarbonate.

Techniques: Recombinant, Gene Expression, Expressing