Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Heat map analysis of the top 20 cis-regulated lncRNAs in the lncRNA high-throughput sequencing and the binding and site results of the NKILA promoter region predicted by JASPAR database. (A) Heatmap demonstrating the top 20 cis-regulated lncRNAs in the control group compared with the normal group. ‘Normal’ indicates blank groups and ‘control’ indicates TGF-β1 (10 ng/ml) induced HK-2 cells. Y-axis shows cis-regulated lncRNA corresponding names. Color changes represent the expression level of the control group changes after normalization treatment compared with the blank group. The change from blue to red after normalization ranges from 0–2. (B) STAT3 binding site information predicted by the JASPAR database for the promoter region from 2000-99 bp upstream of NKILA. lncRNA, long non-coding RNA.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Next-Generation Sequencing, Binding Assay, Control, Expressing

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: lncRNA NKILA overexpression lentivirus and negative control lentivirus transfection experiments. (A) Relative expression level of NKILA after transfection with Lv-NKILA detected by RT-qPCR (n=3). (B) Relative expression levels of NKILA after transfection with Lv-NKILA-shRNA and Lv-NKILA-shRNA (n=3) detected by RT-qPCR. LVCON313 indicates transfection of the knockdown negative control virus LVCON313. LVCON335 indicates transfection of the overexpressed negative control virus LVCON335. Lv-NKILA (76304) indicates transfection of overexpressing Lv-NKILA (76304) in normally cultured HK-2 cells (MOI=5). Lv-NKILA-shRNA (108126) indicates transfection of the knockdown virus Lv-NKILA-shRNA (108126) in normally cultured HK-2 cells (MOI=5). Lv-NKILA-shRNA (108127) indicates transfection of the knockdown virus Lv-NKILA-shRNA (108127) in normally cultured HK-2 cells (MOI=5). ## P<0.01 compared with the normal group. RT-qPCR, reverse transcription quantitative PCR; MOI, multiplicity of infection; shRNA, short hairpin; Lv, lentivirus.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Over Expression, Negative Control, Transfection, Expressing, Quantitative RT-PCR, shRNA, Knockdown, Virus, Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Infection

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Partial EMT phenotype proteins in HK-2 cells transfected with lncRNA NKILA overexpression virus (A) Representative western blotting images and (B) quantification of EMT phenotypic indicators. (C) Statistical analysis results of E-cad fluorescence intensity (n=3). (D) RT-qPCR statistical results of EMT phenotype indicators (n=3). ‘Normal’ indicates after starvation treatment, HK-2 cells were replaced with fresh complete medium and continued to be cultured for 24 h. Control, following starvation treatment, 10 ng/ml TGF-β1 cytokine diluent was added to induce HK-2 cells to construct the renal interstitial fibrosis model for 24 h. ‘OE-NKILA’ indicates after starvation, overexpressed Lv-NKILA (76304) was used for transfection and cells were collected after 24 h of lentivirus transfection. Compared with OE-NC ## P<0.01. E-cad; epithelial-cadherin; RT-qPCR, reverse transcription quantitative PCR; EMT, epithelial-mesenchymal transition; NC, negative control; Lv, lentivirus; OE, overexpression; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin; ns, not significant.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Transfection, Over Expression, Virus, Western Blot, Fluorescence, Quantitative RT-PCR, Cell Culture, Control, Construct, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JKA2/STAT3 pathway protein detection in HK-2 cells transfected with LncRNA NKILA overexpression virus (A) Representative western blotting bands and (B) semi-quantification of results of protein expression of phosphorylated JAK2 and STAT3 (n=3). (C) RT-qPCR statistical results (n=3). ## P<0.01 and # P<0.05 compared with the normal group. ns, not significant; p-, phosphorylated; OE, overexpression; NC, negative control; JAK, Janus kinase; RT-qPCR, reverse transcription quantitative PCR.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Transfection, Over Expression, Virus, Western Blot, Expressing, Quantitative RT-PCR, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Partial EMT phenotype proteins in HK-2 cells transfected with lncRNA NKILA knockdown lentivirus (A) Representative western blotting bands and (B) semi-quantification results of EMT phenotypic indicators. (C) E-cad fluorescence intensity and (D) EMT phenotypic index RT-qPCR (n=3). ‘Normal’ indicates starvation treatment. Compared with normal group ## P<0.01. Compared with control + KD-NC group, **P<0.01 and *P<0.05. EMT, epithelial-mesenchymal transition; E-cad, epithelial-cadherin; RT-qPCR, reverse transcription quantitative PCR; KD, knockdown; Lv, lentivirus; shRNA, short hairpin RNA; NC, negative control; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Transfection, Knockdown, Western Blot, Fluorescence, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, shRNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 signaling pathway-associated proteins in HK-2 cells transfected with LncRNA NKILA knockdown lentivirus (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Statistical analysis results of reverse transcription-quantitative PCR for channel indicators. ## P<0.01 compared with the normal group (n=3), **P<0.01 and *P<0.05 compared with the control + KD-NC group. JAK, Janus kinase; KD, knockdown; NC, negative control; p-, phosphorylated.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Transfection, Knockdown, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 pathway protein detection in the recovery experiment of HK-2 cells treated with AG490. (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Reverse transcription-quantitative PCR for channel indicators (n=3). ## P<0.01 compared with the normal group, **P<0.01 and *P<0.05 compared with control + DMSO group and △△ P<0.01 compared with OE-NKILA group. OE, overexpression; p-, phosphorylated; JAK, Janus kinase; ns, not significant.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Over Expression

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Detection of EMT-related phenotypic proteins in the AG490 intervention HK-2 cell recovery experiment (A) Representative western blotting bands and (B) semi-quantification of EMT phenotypic protein changes (n=3). (C) Statistical analysis results of E-cad fluorescence intensity (n=3) and (D) EMT phenotypic index through reverse transcription quantitative PCR (n=3). ‘Normal’ indicates starvation treatment, HK-2 cells were replaced with fresh complete medium and continued to be cultured for 24 h. ## P<0.01 compared with the normal group, ** P<0.01 and *P<0.05 compared with the control + DMSO group and △△ P<0.01 and △ P<0.05 compared with the OE-NKILA group. EMT, epithelial-mesenchymal transition; E-cad, epithelial cadherin; OE, overexpression; Lv, lentivirus; FN, fibronectin; Col1, collagen I; α-SMA, α-smooth muscle actin; Vim; vimentin.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Cell Recovery, Western Blot, Fluorescence, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Culture, Control, Over Expression

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Immunofluorescence detection of E-CAD protein in the recovery experiment of HK-2 cells treated with AG490. E-cad immunofluorescence staining (magnification, ×400; scale bar, 20 µm; n=3) compared with the normal group. E-cad, epithelial cadherin; OE, overexpression.

Article Snippet: HK-2 cells (cat. no. CL-0109) were cultured in RPMI 1640 medium (cat. no. L220KJ; Shanghai Basal Media Technologies Co., Ltd), supplemented with 10% FBS (cat. no. FBSSR-01021-5, Suzhou Cyagen Biosciences Inc.) at 37°C in a humified 5% CO 2 atmosphere in a CO 2 incubator.

Techniques: Immunofluorescence, Staining, Over Expression