Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: Expression and localization of Snail1 protein in SNAI1 -overexpressed renal tubular cells. (A) Snail1 protein expression was evaluated in vector-control and SNAI1 -overexpressed HK-2 cells by immunoblotting. (B) The Snail1 band intensity was quantified and normalized by that of GAPDH (loading control). (C) Expression and localization of Snail1 protein were also examined by immunofluorescence assay. (D) The Snail1 fluorescence intensity was evaluated in at least 100 cells across 10 random fields per sample. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Expressing, Plasmid Preparation, Control, Western Blot, Immunofluorescence, Fluorescence, Derivative Assay

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: Validation of differentially expressed proteins in SNAI1 -overexpressed renal tubular cells. (A, B) The significantly decreased and increased proteins in SNAI1 -overexpressed HK-2 cells were validated by immunoblotting compared with vector-control HK-2 cells. (C – E) Their band intensities were quantified and normalized by that of β-actin (loading control). Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Biomarker Discovery, Western Blot, Plasmid Preparation, Control, Derivative Assay

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression up-regulated nucleophosmin expression in renal tubular cells. (A) Expression and localization of nucleophosmin in vector-control and SNAI1 -overexpressed HK-2 cells were examined by immunofluorescence assay. (B) The nucleophosmin fluorescence intensity was evaluated in at least 100 cells across 10 random fields per sample. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Expressing, Plasmid Preparation, Control, Immunofluorescence, Fluorescence, Derivative Assay

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression increased nucleolar organizer regions (NORs) in renal tubular cells. ( A ) The NORs in vector-control and SNAI1 -overexpressed HK-2 cells were evaluated by AgNOR staining. The inset indicates the zoom-in image of the highlighted area. (B, C) The NORs were quantified in individual nuclei from at least 15 random fields per sample based on (B) their total area per nucleus and (C) their number per nucleus. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Plasmid Preparation, Control, Staining, Derivative Assay

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression induced cell enlargement and increased granularity of renal tubular cells. (A) The vector-control and SNAI1 -overexpressed HK-2 cells were imaged under a phase contrast microscope. (B) Cell areas were measured from at least 100 cells in 10 random fields per sample. (C) The cells were also analyzed by flow cytometry. Representative scatter plots show forward scatter (FSC; indicating cell size) on the x -axis and side scatter (SSC; indicating cell granularity) on the y -axis. (D) The overlaid histogram plot displays a shift in cell granularity (SSC) of SNAI1 -overexpressed HK-2 cells compared with vector-control cells. (E, F) Cell size (E) and cell granularity (F) were quantified and averaged from at least 10,000 cells per sample. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Plasmid Preparation, Control, Microscopy, Flow Cytometry, Derivative Assay

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression increased senescence markers in renal tubular cells. (A) Levels of senescence markers, including p21 and γH2AX, in vector-control and SNAI1 -overexpressed HK-2 cells were evaluated by immunoblotting. (B, C) Their band intensities were quantified and normalized by that of β-actin (loading control). Each bar shows the mean ± standard error of the mean derived from three independent experiments. (D) Expression of γH2AX was also evaluated by immunofluorescence assay. (E) Magnified images revealed the formation of γH2AX-foci within the nuclei as depicted by vivid green spots. The red line indicates the direction for fluorescence intensity spectral profiling. (F) The fluorescence intensity spectral profiles of γH2AX expression (illustrated by the green line) and nuclei (illustrated by the blue line).

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Plasmid Preparation, Control, Western Blot, Derivative Assay, Expressing, Immunofluorescence, Fluorescence

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression increased the production of matrix metalloproteinase (MMP) from renal tubular cells. (A) MMP-9 in culture supernatants derived from vector-control and SNAI1 -overexpressed HK-2 cells was evaluated by gelatin zymography. (B) MMP-9 band intensity was quantified. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Derivative Assay, Plasmid Preparation, Control, Zymography

Journal: Genes & Diseases

Article Title: Deciphering the impact of SNAI1 gene on renal tubular cell proteome, nucleolar stress, ribosome biogenesis, senescence, DNA damage response, and focal adhesion dynamics

doi: 10.1016/j.gendis.2025.101926

Figure Lengend Snippet: SNAI1 overexpression induced focal adhesion organization in renal tubular cells. (A) Paxillin expression was evaluated in vector-control and SNAI1 -overexpressed HK-2 cells by immunoblotting. (B) The paxillin band intensity was quantified and normalized by that of β-actin (loading control). (C) Expression and localization of paxillin were also examined by immunofluorescence assay. (D) The paxillin fluorescence intensity was evaluated in at least 100 cells across 10 random fields per sample. Each bar shows the mean ± standard error of the mean derived from three independent experiments.

Article Snippet: Following 48 h, the supernatant containing released viral particles was collected and filtered through a 0.45-μm syringe filter before being introduced to HK-2 cells (ATCC) (the target cells set at 40% confluence).

Techniques: Over Expression, Expressing, Plasmid Preparation, Control, Western Blot, Immunofluorescence, Fluorescence, Derivative Assay

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: Human Mutation

Article Title: Cross‐Cohort Transcriptomic Integration Identifies IFIT2 as a Translational Diagnostic Biomarker and Functional Driver of Inflammation‐Linked Tubular Injury in Chronic Kidney Disease

doi: 10.1155/humu/8282277

Figure Lengend Snippet: Induction and knockdown of IFIT2 in renal tubular epithelial cells. (A–B) IFN‐ γ –induced IFIT2 expression in HK‐2 and RPTEC cells. (C–D) TGF‐ β 1–induced IFIT2 expression in HK‐2 and RPTEC cells. (E–F) Validation of IFIT2 knockdown efficiency by qPCR. Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Human renal proximal tubular epithelial cells, including the HK‐2 cell line (ATCC, RRID: CVCL_0302) and primary RPTEC cells (ATCC, RRID: CVCL_K278), were used in this study.

Techniques: Knockdown, Expressing, Biomarker Discovery

Journal: Human Mutation

Article Title: Cross‐Cohort Transcriptomic Integration Identifies IFIT2 as a Translational Diagnostic Biomarker and Functional Driver of Inflammation‐Linked Tubular Injury in Chronic Kidney Disease

doi: 10.1155/humu/8282277

Figure Lengend Snippet: IFIT2 knockdown attenuates IFN‐ γ –induced injury and apoptosis in renal tubular epithelial cells. (A–B) CCK‐8 assay showing that IFIT2 knockdown alleviates IFN‐ γ –induced reduction of cell viability in HK‐2 and RPTEC cells. (C–F) Annexin V/PI flow cytometry analysis showing that IFIT2 knockdown reduces IFN‐ γ –induced apoptosis in (C, E) HK‐2 and (D, F) RPTEC cells. Data are presented as mean ± SD from three independent experiments. ∗∗∗ p < 0.001.

Article Snippet: Human renal proximal tubular epithelial cells, including the HK‐2 cell line (ATCC, RRID: CVCL_0302) and primary RPTEC cells (ATCC, RRID: CVCL_K278), were used in this study.

Techniques: Knockdown, CCK-8 Assay, Flow Cytometry