Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: Identification of lncRNA LINC02470 as a negative regulator of NK cytotoxicity. (A) Identification of NK cell regulator LINC02470 . (B) LINC02470 expression by RT-PCR. VPA-untreated and treated HCC NK cells, VPA-untreated and treated NK-92MI cells were collected for RT-PCR. A representative agarose gel electrophoresis is shown. (C) Quantitation of LINC02470 expression by RT-qPCR in NK cells from HCC patients (n=10) and NK-92MI cells untreated and treated by VPA. ***p<0.001. (D) Negative association between LINC02470 expression and NK cytotoxicity in NK cells from healthy donors (n=10) and HCC patients (n=10). ***p<0.001.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Quantitation Assay, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 knockdown increased (A–C) , and LINC02470 overexpression suppressed (D–F) the cytotoxicity of NK-92MI cells. shCT: NK-92MI transfected with random control shRNA vector; shLINC02470: NK-92MI transfected with shLINC02470 vector. CT: NK-92MI transfected with blank control plasmid. LINC02470 -OE: NK-92MI transfected with LINC02470 overexpression plasmid. Data represent ten independent experiments. ***p<0.001. **p<0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Knockdown, Over Expression, Transfection, Control, shRNA, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: Location of LINC02470 in NK-92MI cells. (A) LINC02470 location assays by cytoplasmic and nuclear separation and PCR. U6: nuclear control. β-Actin: cytoplasm control. (B) Confirmation of LINC02470 location by RNA-FISH. DAPI: nuclear control. Both results showed that LINC02470 was primarily located in the nucleus.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 suppressed the expression of NCR1 . (A) Schematic diagram of gene NCR1 structure. 5’-Enh, 3’-Enh: the NCR1 5’- and 3’-enhancers; p NCR1 : NCR1 promoter; E1-E7: NCR1 exons. 5’-CT, 3’-CT: the RAT control sites. (B) Primers were designed in different regions of NCR1 gene, and the RAT pulldown complex was used to map the LINC02470 binding by qPCR. Note the enrichment of the LINC02470 binding signals in the 5’-enhancer and promoter region (5’-Enh, Prot-1 and Prot-2) (***P < 0.001, *P < 0.05 as compared with the RAT control). Data represent four independent experiments. (C) The relative expression of NCR1 was detected by qPCR in NK-92MI, shCT and shLINC02470 cells, respectively. The results showed that NCR1 expression significantly increased after LINC02470 knockdown. Experiments were performed in triplicate. ***P < 0.001. (D) Representative flow cytometry analysis of NKp46 expression in NK-92MI and shLINC02470 cells. Both percentage and mean fluorescence intensity (MFI) of NKp46 expression increased after knocking down LINC02470 . Three independent experiments were performed. **P < 0.01. (E) The role of NKp46 in NK cell cytotoxicity. Both NK-92MI cells and shLINC02470 cells were pre-incubated with anti-NKp46 antibody and were then tested for killing of K652 cells. Data were obtained from ten independent experiments. NK-92MI-CT: NK-92MI cells pre-incubated with an isotype control IgG; NK-92MI-BL: NK-92MI cells pre-incubated with anti-NKp46 antibody; shLINC02470: NK-92MI transfected with shLINC02470 vector; shLINC02470-CT: shLINC02470 cells pre-incubated with an isotype control IgG; shLINC02470-BL: shLINC02470 cells pre-incubated with anti-NKp46 antibody. **P < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Expressing, Control, Binding Assay, Knockdown, Flow Cytometry, Fluorescence, Incubation, Transfection, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 damaged the maintenance of intrachromosomal looping in the NCR1 locus. (A) The PCR sites in the NCR1 locus. 5’-Enh: 5’-enhancer; p NCR1 : NCR1 promoter; E1-E7: Exons; 3’-Enh: 3’-Enhancer. 5’-CT, 3’-CT: the RAT control sites. Arrows: intrachromosomal interactions. (B) Intrachromosomal interactions were quantitated by 3C (chromatin conformation capture) qPCR. Knockdown of LINC02470 raised intrachromosomal interaction loops. Data were obtained from five independent experiments. For comparison, the relative 3C interaction was calculated by setting the 5’- and 3’- controls as 1 (**P < 0.01 as compared with the NK-92MI controls). (C) Sequencing of the NCR1 intrachromosomal loop products. Arrows: the 3C ligation product containing the MboI site that is flanked by the promoter and the enhancer sequences.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Control, Knockdown, Comparison, Sequencing, Ligation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 inhibits the NCR1 enhancer RNA pathway. (A) Location of NCR1 enhancer RNAs. (B) LINC02470 knockdown activates NCR1 eRNA in 5’ enhancer. Expression of NCR1 eRNAs in NK-92MI, shCT and shLINC02470 cells was measured by RT-qPCR. shCT: NK-92MI transfected with the random shRNA vector control; shLINC02470: NK-92MI transfected with shLINC02470. Gene expression was normalized to β-Actin control. All experiments were performed in triplicate and statistically significant differences by Student’s t test. **P < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Transfection, shRNA, Plasmid Preparation, Control, Gene Expression

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 abolished enrichments of NCR1 DNA sequences for H3K4me3, but enhanced H3K9me3 in the promoter of NCR1 gene. Two pairs of primers were designed in the promoter of NCR1 gene. Enrichments of NCR1 DNA sequences for H3K4me3, H3K9me3 occupancy in NK-92MI and shLINC02470 cells were measured using ChIP-qPCR. Data were obtained from three independent experiments. **p < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: ChIP-qPCR

Journal: AMB Express

Article Title: Myeloperoxidase-DNA complex: a marker and combined target for Pseudomonas aeruginosa -associated bronchiectasis

doi: 10.1186/s13568-026-02012-w

Figure Lengend Snippet: Effects of NETs intervention on viability, inflammation and oxidative stress levels of BEAS-2B cells. A CCK-8 assay and microscopic observation under a 10x objective. Comparison of ROS ( B ), MDA ( C ), IL1β ( D ) and IL6 ( E ) in BEAS-2B cells between NETs group and control group ( n = 3, P < 0.05)

Article Snippet: To evaluate the effect of NETs stimulation on airway epithelial cells, subsequent experiments were performed using the human bronchial epithelial cell line BEAS-2B (Procell, Wuhan, China).

Techniques: CCK-8 Assay, Comparison, Control

Journal: Hereditas

Article Title: RBM15 promotes COAD progression by regulating the m6A modification of TMC5

doi: 10.1186/s41065-025-00530-4

Figure Lengend Snippet: Expression level of TMC5 in COAD. ( A ) Analysis of mRNA expression of TMC5 in different cancers based on TIMER2.0 database. ( B ) GEPIA database ( http://gepia.cancer-pku.cn/ ) exhibited the expression level of TMC5 in COAD samples ( n = 275) and normal samples ( n = 349). ( C and D ) UALCAN database showed TMC5 mRNA and protein expression based on The Cancer Genome Atlas (TCGA) and Clinical Proteomic Tumor Analysis Consortium (CPTAC) samples. ( E ) TMC5 protein level was determined in 5 normal tissues and 5 COAD tumor tissues using western blot. ( F ) TMC5 mRNA level was detected in 69 normal tissues and 69 COAD tumor tissues using RT-qPCR. ( G ) Immunohistochemical observation of TMC5 expression in normal and COAD tumor tissues. ( H ) Western blot analysis of TMC5 protein level in NCM460 cell line and COAD cell lines (Caco-2, SW620, and LoVo). *** P < 0.001

Article Snippet: In an incubator at 37 ̊C with 5% CO 2 , COAD cell lines (Caco-2, CL-0050; SW620, CL-0225; LoVo, CL-0144; Procell, Wuhan, China) were maintained in corresponding media (CM-0050, CM-0225, CM-0144; Procell).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemical staining

Journal: Hereditas

Article Title: RBM15 promotes COAD progression by regulating the m6A modification of TMC5

doi: 10.1186/s41065-025-00530-4

Figure Lengend Snippet: TMC5 downregulation repressed COAD cell development. Caco-2 and LoVo cells were transfected with sh-NC or sh-TMC5. ( A ) TMC5 protein level was determined in transfected Caco-2 and LoVo cells using western blot. ( B ) Cell proliferation was assessed using EdU assay. ( C ) Cell apoptosis was measured using flow cytometry assay. ( D ) Caspase 3 activity was detected using a commercial kit. ( E and F ) Cell migration and invasion were examined using Transwell assays. ( G ) ROS level was examined using a commercial kit. ( H ) Iron assay kit was carried out to detect Fe + level in transfected Caco-2 and LoVo cells. ( I and J ) FTH1 and xCT mRNA levels were determined using RT-qPCR. ** P < 0.01, *** P < 0.001

Article Snippet: In an incubator at 37 ̊C with 5% CO 2 , COAD cell lines (Caco-2, CL-0050; SW620, CL-0225; LoVo, CL-0144; Procell, Wuhan, China) were maintained in corresponding media (CM-0050, CM-0225, CM-0144; Procell).

Techniques: Transfection, Western Blot, EdU Assay, Flow Cytometry, Activity Assay, Migration, Iron Assay, Quantitative RT-PCR

Journal: Hereditas

Article Title: RBM15 promotes COAD progression by regulating the m6A modification of TMC5

doi: 10.1186/s41065-025-00530-4

Figure Lengend Snippet: Inhibiting TMC5 impaired COAD cell growth in vivo. sh-NC or sh-TMC5-transfected LoVo cells were subcutaneously injected into mice. ( A and B ) Tumor volume and tumor weight were examined. ( C ) IHC staining was used to assess the positive expression of TMC5, ki-67, and FTH1 in xenografted tumor samples. ** P < 0.01, *** P < 0.001

Article Snippet: In an incubator at 37 ̊C with 5% CO 2 , COAD cell lines (Caco-2, CL-0050; SW620, CL-0225; LoVo, CL-0144; Procell, Wuhan, China) were maintained in corresponding media (CM-0050, CM-0225, CM-0144; Procell).

Techniques: In Vivo, Transfection, Injection, Immunohistochemistry, Expressing

Journal: Hereditas

Article Title: RBM15 promotes COAD progression by regulating the m6A modification of TMC5

doi: 10.1186/s41065-025-00530-4

Figure Lengend Snippet: RBM15 modulated m6A enrichment and stability of TMC5 mRNA. ( A ) The online tool SRAMP was used to predict m6A sites of TMC5. ( B ) Alterations in the m6A methylation level of TMC5 after inhibition of RBM15 were analyzed by MeRIP-qPCR assay. ( C ) Influences of RBM15 overexpression or downregulation on TMC5 mRNA stability after Actinomycin D treatment was measured using RT-qPCR in Caco-2 and LoVo cells. ( D ) RBM15 and TMC5 protein levels were determined in Caco-2 and LoVo cells transfected with vector, RBM15, sh-NC, or sh-RBM15 using western blot. ( E ) Their interaction was confirmed using dual-luciferase reporter assay in Caco-2 and LoVo cells. ( F ) UALCAN database presented the expression level of RBM15 in COAD samples ( n = 286) and normal samples ( n = 41) based on sample types TCGA samples. ( G ) RT-qPCR analysis of RBM15 mRNA level in 69 normal tissues and 69 COAD tumor tissues. ( H ) Western blot analysis of RBM15 protein level in 5 normal tissues and 5 COAD tumor tissues. ( I ) IHC staining analyzed the positive expression rate RBM15 in normal tissues and COAD tumor tissues. ( J ) GEPIA database exhibited the association between RBM15 and TMC5 expression in COAD samples. ( K ) Pearson correlation analysis was applied to evaluate the expression correlation between RBM15 and TMC5 in COAD tissues. ( L ) RBM15 protein level was detected in NCM460 cell line and COAD cell lines (Caco-2 and LoVo) using western blot. ** P < 0.01, *** P < 0.001

Article Snippet: In an incubator at 37 ̊C with 5% CO 2 , COAD cell lines (Caco-2, CL-0050; SW620, CL-0225; LoVo, CL-0144; Procell, Wuhan, China) were maintained in corresponding media (CM-0050, CM-0225, CM-0144; Procell).

Techniques: Methylation, Inhibition, Over Expression, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Luciferase, Reporter Assay, Expressing, Immunohistochemistry

Journal: Hereditas

Article Title: RBM15 promotes COAD progression by regulating the m6A modification of TMC5

doi: 10.1186/s41065-025-00530-4

Figure Lengend Snippet: RBM15/TMC5 regulated COAD cell malignant behaviors. ( A ) Western blot analysis of TMC5 protein level in Caco-2 and LoVo cells transfected with vector or TMC5. ( B-K ) Caco-2 and LoVo cells were transfected with sh-NC + vector, sh-RBM15 + vector, or sh-RBM15 + TMC5. ( B ) TMC5 protein level was monitored by western blot. ( C and D ) Cell proliferation and apoptosis were tested using EdU and flow cytometry assays. ( E ) Caspase 3 activity was examined using a commercial kit. ( F and G ) Cell migration and invasion were measured using Transwell assays. ( H and I ) ROS level and Fe + level were determined by special kits. ( J and K ) FTH1 and xCT mRNA levels were examined using RT-qPCR. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: In an incubator at 37 ̊C with 5% CO 2 , COAD cell lines (Caco-2, CL-0050; SW620, CL-0225; LoVo, CL-0144; Procell, Wuhan, China) were maintained in corresponding media (CM-0050, CM-0225, CM-0144; Procell).

Techniques: Western Blot, Transfection, Plasmid Preparation, Flow Cytometry, Activity Assay, Migration, Quantitative RT-PCR

Journal: Advanced Science

Article Title: PACS‐2 Mitigates NPSC Apoptosis and Intervertebral Disc Degeneration by Preserving MAM Integrity via the SP1/LRRK2/Mfn2 Axis

doi: 10.1002/advs.202511781

Figure Lengend Snippet: Impaired PACS‐2 expression and MAM integrity in degenerative NP tissues of humans and rats, as well as acid‐treated NPSCs. A) Representative MRI images, General views (Scale bar: 1 cm), HE staining images(Scale bar: 25 µm), and Alcian blue staining images (Scale bar: 25 µm) of human IVD tissues with Grade II and Grade IV degeneration. B,C) Representative immunohistochemical staining images of PACS‐2 in human NP tissues and the quantitative analysis (n = 5). Scale bar: 25 µm. D,E) Representative western blot images and quantitative analysis of PACS‐2 in human NP tissues (n = 5). F) Representative immunofluorescence double staining images of IP3R1 and VDAC1 in human NP tissues. Scale bar: 20 µm. G,H) Representative immunohistochemical staining images of Cleaved caspase‐3 in human NP tissues and the quantitative analysis (n = 5). Scale bar: 25 µm. I) Representative MRI images of rat tails. J) Representative HE staining images and SO staining images of rat IVD tissues. Scale bar: left panel, 500 µm; right panel, 100 µm. K,L) Representative immunohistochemical staining images of PACS‐2 in rat NP tissues and the quantitative analysis (n=5). Scale bar: 25 µm. M,N) Representative western blot images and quantitative analysis of PACS‐2 in rat NP tissues (n = 5). O) Representative immunofluorescence double staining images of IP3R1 and VDAC1 in rat NP tissues. Scale bar: 5 µm. P,Q) Representative immunohistochemical staining images of Cleaved caspase‐3 in rat NP tissues and the quantitative analysis (n=5). Scale bar: 25 µm. R) UMAP visualization of cell populations distribution between mild IDD (Pfirrmann II/III) group and severe IDD (Pfirrmann IV/V) group. S) Percentage of different cell types in mild and severe IDD group. T,U) Representative western blot images and quantitative analysis of PACS‐2 and Cleaved caspase‐3 in human NPSCs exposed to acid medium of pH 6.5 for 24 h (n = 3). V) Representative immunofluorescence staining images of PACS‐2 in the acid‐treated NPSCs (n=3). Scale bar: 10 µm. W) Representative fluorescence confocal images with Mito‐Tracker (green) and ER‐Tracker (red) staining in the acid‐treated NPSCs. Scale bar: 10 µm. Representative TEM images of NPSCs. Scale bar: 200 nm. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Alizarin Red (Procell, Wuhan, China), Oil Red O (Procell, Wuhan, China), and Alcian Blue (Procell, Wuhan, China) staining were performed as instructed by the manufacturer.

Techniques: Expressing, Staining, Immunohistochemical staining, Western Blot, Immunofluorescence, Double Staining, Fluorescence