sipklr origen  (OriGene)

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) UMAP projection of all endothelial cells from Tsukui et al.(22) (light blue-healthy red-IPF)(4,338 cells) (B) Volcano plot comparing genes differentially expressed across healthy and IPF endothelial cells. (C) UMAP projection of all endothelial cells from Tsukui et al. and proportion analysis (D) Heatmap of immune, hypoxic and cytoskeletal genes enriched in ACKR1pos VECs. (E) UMAP projection with gene expression for ACKR1 and COL15A1 split between healthy and IPF cells. (F) IF for ACKR1 and SELP in human IPF precision cut lung slices (large scale 50µm, small scale 20µm) (G) IF for ACKR1 and VCAM1 in human IPF precision cut lung slices (large scale 50µm, small scale 10µm) (H) IF for ACKR1 and HIF1A in healthy and IPF lungs (large scale 50µm, small scale 5µm). (I) Schematic for precision cut lung slices (F) IF for ACKR1, CD45 and COL1A1 in human IPF precision cut lung slices (large scale 50µm, small scale 20µm)(FF=Fibroblastic Foci). (K) IF for ACKR1 and aSMA in healthy and IPF lungs (large scale 50µm)

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Gene Expression

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) UMAP projection of all endothelial cell markers from Tsukui et al. (B) Differentially expressed genes between all endothelial subpopulations. (C) Violin plot with ACKR1 normalized expression in healthy and IPF endothelial cells. (D) Violin plot with ACKR1 normalized expression across all lung lineages. (E) GO enrichment of upregulated and downregulated genes.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Expressing

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Circle plot showing CellChat analysis of outgoing and incoming signaling in IPF lungs and (B) heatmap depicting relative interaction strength between senders and receivers. (line thickness indicates relative communication probability). (C) Enrichment and spatial mapping of the ACKR1 VECs gene signature in human lung spatial transcriptomics data (Franzén et al.(29)). (D) Heatmap displaying the mean spatial proximity scores between source endothelial populations CPE+ / CDH5+ double-positive spots in Healthy Controls (left) and ACKR1 signature-scoring spots in IPF(right) and various target cell types. (E) Enrichment of the ACKR1 VEC-CTHRC1 gene signature across healthy and IPF samples from the Franzén et al (29) dataset and proximity quantification. F) Enrichment of the ACKR1 VEC-CD14 gene signature across healthy and IPF samples from the Franzén et al (29) dataset and proximity quantification. (G) IF for ACKR1 in an IPF lung (large scale 500µm, small scale 50µm). (H) HE, ACKR1 VEC signature enrichment and fibrosis score enrichment on mild and severe IPF biopsy spatial RNA. Statistical significance: (C,E,F) non-parametric Wilcoxon rank-sum test; (D) two-sided Mann–Whitney U test.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Spatial Transcriptomics, MANN-WHITNEY

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Spatial RNA sequencing experimental set up. (B) Bar plot of average gene content of spatial RNA sequencing samples. (C) Bar plot of spot count of spatial RNA sequencing samples. ( D ) Spatial mapping and quantification of proximity between ACKR1 VECs and ligands in healthy and IPF lungs from the Franzén et al (29) dataset. Statistical significance was evaluated using a non-parametric Wilcoxon rank-sum test.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Circle plot depicting CellChat analysis of outgoing and incoming signaling in IPF lungs and heatmap showing relative interaction strength between senders and receivers (line thickness indicates relative communication probability). (B) Bubble plot with Ligand-Receptor pair interactions between ACKR1to macrophages (left), Macrophages to ACKR1 VECs and Monocytes to ACKR1 VECs. Color indicates normalized interaction strength. (C) IF for ACKR1 and CD68 in IPF precision cut lung slices (scale 50µm). (D) IF for ACKR1 and CCR5 in IPF precision cut lung slices (scale 50µm). (E) IF for ACKR1 and SPP1 in IPF precision cut lung slices (scale 50µm).

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques:

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Schematic for ACKR1 + VEC isolation (B) Brightfield image of ACKR1 + and ACKR1 - ECs. (scale 125 pixels) (C) Western blot for ACKR1 (D) Boxplots showing normalized mRNA expression for inflammatory and hypoxic markers (E) Schematic for conditional media experimental set up ( F) Boxplots showing normalized mRNA expression for CTHRC1, COL1A1 and ACTA2 (G) Schematic for immune cell adhesion experimental set up (H) Fluorescently labeled THP1 cells adhered to ACKR1 - and ACKR1 + ECs and quantification of adhered cells (scale 250 pixels). (I) Schematic for immune cell migration experimental set up (J) Quantification of migrated THP1 cells. Statistical analysis: (D,H,J) two-tailed Student’s t-test and (F) a one-way ANOVA.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Isolation, Western Blot, Expressing, Labeling, Migration, Two Tailed Test

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Boxplot showing normalized mRNA expression for ACKR1 (B) Fluorescent labeled THP1 cells adhered to Scramble or ACKR1-siRNA treated ACKR1pos ECs and quantification (scale 250 pixels) (C) Boxplots showing normalized mRNA expression for COL1A1, FN1, CTHRC1, ACTA2 and TNC (D) Schematic for collagen contraction assay experimental set up (E) Representative images of collagen contraction and quantification of collagen area (F) Heatmap for immune recruiting, profibrotic and cytoskeletal genes (G) GO enrichment of upregulated/downregulated genes. (H) IF staining for p65 and ACKR1 in healthy and IPF lungs (top scale 20µm, bottom scale 5µm). (I) Western blot for P-p65 and p65 Statistical significance: Statistical analysis: (B,C) two-tailed Student’s t-test and (A,E) a one-way ANOVA.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Expressing, Labeling, Contraction Assay, Staining, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) IF for ACKR1 and Slc6a2 in bleomycin treated mouse lungs (scale 20 µm). (B) IF for ACKR1 and CD45 in bleomycin treated mouse lungs (large scale 20µm, small scale 10µm). (C) IF for ACKR1 and CD45 in mouse lungs twenty-one days after bleomycin installation (scale 50µm). (D) IF for ACKR1, CD68 and Col1a1 in mouse lungs twenty-one days after bleomycin installation (scale 200 µm). (E) IF for ACKR1, Slc6a2 and Col1a1 in mouse lungs twenty-one days after bleomycin installation (scale 100 µm). (F) scRNAseq set-up and UMAP projection of all venous endothelial cells (purple-sham, orange-seven days post bleomycin administration (cells) (G) UMAP projection of ACKR1 positive and ACKR1 negative cells and proportion plot (H) Heatmap with differentially expressed genes. (I) Bubble plot with inflammatory and immune recruiting genes (G) GO enrichment of upregulated genes. (K) Day seven ACKR1 signature on the Adams et al(11)., Habermann et al.(12) and Tsukui et al. (22) datasets. Statistical significance was evaluated using a two-sided Wilcoxon rank-sum test.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques:

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) UMAP projection of all endothelial cells from sham and day seven post bleomycin and proportion plot. (B) UMAP projection of endothelial cells split between sham (purple) and day seven post bleomycin (orange). (C) Bubble plot with representative markers for each endothelial lineage. (D) Differential gene expression in each endothelial lineage. (E) UMAP plots with normalized gene expression across venous endothelial cells (F) Human and mouse ACKR1 VEC signature enrichment on all endothelial lineages from sham and bleomycin injured mice. (G) KEGG enrichment on mouse ACKR1pos VECs . (H) IF for ACKR1 and SPP1 in day day21 mouse bleomycin treated lungs (scale= µm). (I) IF for ACKR1 and EdU in day day7 mouse bleomycin treated lungs (scale= 10µm).

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Gene Expression

Journal: bioRxiv

Article Title: ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

doi: 10.64898/2026.07.31.742106

Figure Lengend Snippet: (A) Experimental set up. (B) Diagram with representative body weight. (C) Bar plot depicting left lobe wet weight. (D) Bar plot depicting left lobe hydroxyproline measurement (E) Masson’s trichrome staining of sham, carrier and amikacin treated lungs (scale 100 µm). (F) IF for ACKR1, CD45 and CD68 in sham, carrier and amikacin treated lungs (scale 20 µm). (G) IF for ACKR1, aSMA and Col1a1 in sham, carrier and amikacin treated lungs (scale 20 µm). Statistical significance: (C,D) one-way ANOVA.

Article Snippet: IPF-derived ACKR1+ VEC cells were cultured and treated with either scramble siRNA (Horizon Discovery/Dharmacon, D-001810-01-05) or siRNA targeting ACKR1 (MedChemExpress, 5500343490) for two days in OptiMEM media (Fisher, 31985070) for 48 hours, after which the cells were lysed and RNA extracted.Total RNA integrity was verified using RNA 6000 Pico Assay run on an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA).

Techniques: Staining

Journal: iScience

Article Title: Hyaluronic acid and folic acid-modified mesoporous silica nanoparticles delivering sulforaphane suppress NSCLC via SPI1/miR-616-5p axis

doi: 10.1016/j.isci.2026.116293

Figure Lengend Snippet: SPI1 regulates NSCLC cell viability and proliferation (A–C) Western blot analysis of transfection efficiency for SPI1 overexpression and knockdown ( n = 3, n represents biological replicates). (D) CCK-8 assay assessing the effects of SPI1 overexpression and knockdown on NSCLC cell viability ( n = 6, n represents biological replicates). (E) Colony formation assay evaluating the impact of SPI1 overexpression and knockdown on NSCLC cell proliferation ( n = 3, n represents biological replicates). Data are represented as mean ± SD. p values are based on a one-way ANOVA test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Lentiviral shRNA targeting human SPI1 , GeneChem , N/A.

Techniques: Western Blot, Transfection, Over Expression, Knockdown, CCK-8 Assay, Colony Assay

Journal: iScience

Article Title: Hyaluronic acid and folic acid-modified mesoporous silica nanoparticles delivering sulforaphane suppress NSCLC via SPI1/miR-616-5p axis

doi: 10.1016/j.isci.2026.116293

Figure Lengend Snippet: SPI1 regulates NSCLC cell migration, invasion, and EMT (A) Transwell assay assessing the effects of SPI1 on cell migration and invasion ( n = 3, n represents biological replicates), scale bars, 200 μm. (B) Western blot analysis of the impact of SPI1 on the expression levels of EMT-related proteins (N-cadherin, E-cadherin, and vimentin, n = 3, n represents biological replicates). Data are represented as mean ± SD. p values are based on a one-way ANOVA test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Ⅰ, N-cadherin. Ⅱ, E-cadherin. Ⅲ, vimentin. Ⅳ, GAPDH.

Article Snippet: Lentiviral shRNA targeting human SPI1 , GeneChem , N/A.

Techniques: Migration, Transwell Assay, Western Blot, Expressing

Journal: iScience

Article Title: Hyaluronic acid and folic acid-modified mesoporous silica nanoparticles delivering sulforaphane suppress NSCLC via SPI1/miR-616-5p axis

doi: 10.1016/j.isci.2026.116293

Figure Lengend Snippet: SPI1 promotes NSCLC cell invasion and migration by transcriptionally regulating miR-616-5p (A and B) RT-PCR analysis of miR-616-5p expression following SPI1 overexpression or knockdown ( n = 3, n represents biological replicates). (C–E) Transwell assay evaluating cell migration and invasion ( n = 3, n represents biological replicates), scale bars, 200 μm. Data are represented as mean ± SD. p values are based on a one-way ANOVA test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Lentiviral shRNA targeting human SPI1 , GeneChem , N/A.

Techniques: Migration, Reverse Transcription Polymerase Chain Reaction, Expressing, Over Expression, Knockdown, Transwell Assay

Journal: iScience

Article Title: Hyaluronic acid and folic acid-modified mesoporous silica nanoparticles delivering sulforaphane suppress NSCLC via SPI1/miR-616-5p axis

doi: 10.1016/j.isci.2026.116293

Figure Lengend Snippet: SF downregulates miR-616-5p activity by directly binding to and inhibiting SPI1 (A) Molecular docking analysis of SF with SPI1. (B) SPRi binding curve of SF to SPI1. (C and D) Western blot analysis of SPI1 expression in cells ( n = 3, n represents biological replicates). (E) RT-PCR analysis of miR-616-5p expression in cells ( n = 3, n represents biological replicates). Data are represented as mean ± SD. p values are based on a one-way ANOVA test. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Lentiviral shRNA targeting human SPI1 , GeneChem , N/A.

Techniques: Activity Assay, Binding Assay, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Hyaluronic acid and folic acid-modified mesoporous silica nanoparticles delivering sulforaphane suppress NSCLC via SPI1/miR-616-5p axis

doi: 10.1016/j.isci.2026.116293

Figure Lengend Snippet: In vivo targeting and anti-NSCLC efficacy of MSNs@SF-HA-FA (A) In vivo fluorescence imaging analysis ( n = 5, n represents the number of mice). (B) Ex vivo fluorescence imaging of major organs ( n = 5, n represents the number of mice). (C) Images of lung tumor tissues from mice ( n = 5, n represents the number of mice). (D) H&E-stained images and immunohistochemical analysis of SPI1 expression in xenograft tumors ( n = 5, n represents the number of mice), scale bars, 100 μm.

Article Snippet: Lentiviral shRNA targeting human SPI1 , GeneChem , N/A.

Techniques: In Vivo, Fluorescence, Imaging, Ex Vivo, Staining, Immunohistochemical staining, Expressing

Journal: Molecular Therapy Oncology

Article Title: CBX6 and CA9 as predictive indicators and therapeutic targets in GBM

doi: 10.1016/j.omton.2026.201159

Figure Lengend Snippet: CBX6 expression is downregulated in multiple cancer types (A) Statistical analysis of The Cancer Genome Atlas (TCGA) Pan-Cancer RNA sequencing (RNA-seq) database shows CBX6 expression is downregulated in various cancer types. (B) CBX6 expression is significantly suppressed in brain cancer, breast cancer, lung adenocarcinoma (abbreviated as LUAD), and prostate cancer, based on analysis of TCGA tumor microarray data using the BRowse All Variants Online (BRAVO) database method. (C and D) Comparison of CBX6 expression in patient-derived GBM tissues and primary tumor cells relative to respective normal controls using qRT-PCR. GAPDH was used as a loading control. (D) Reduced CBX6 expression is significantly associated with poor prognosis in patients with glioma, as shown in Kaplan-Meier survival curves derived from a public patient-derived microarray database, p = 1.94∗ e−7. PBT, primary brain tumor; RBT, recurrent brain tumor.

Article Snippet: A pCMV6-entry plasmid containing Myc-DDK (same as Flag)-tagged human CBX6 cDNA (Cat#: RC204166) and a CBX6 Human shRNA Plasmid Kit (Cat#: TF314170) were purchased from OriGene.

Techniques: Expressing, RNA Sequencing, Microarray, Comparison, Derivative Assay, Quantitative RT-PCR, Control

Journal: Molecular Therapy Oncology

Article Title: CBX6 and CA9 as predictive indicators and therapeutic targets in GBM

doi: 10.1016/j.omton.2026.201159

Figure Lengend Snippet: CBX6 modulation affects tumor cell morphology and proliferation (A) CBX6 modulation affects the morphology of U-251 MG cells in two-dimensional culture. (B) Downregulation of CBX6 increases growth and invasion of U-251 MG cells in 3D culture. (C) Overexpression of CBX6 significantly suppresses U-251 MG proliferation on days 3 and 5 post-plating, as measured by MTT assay. (D) MTT assay of KLuc cells stably overexpressing human CBX6 or transfected with control plasmid on days 3 and 5 post-plating. (E) Silencing CBX6 increases U-251 MG proliferation compared to shRNA controls. Images were captured randomly from different fields under 20× magnification. VC, vector control; OE, CBX6 overexpression; KD, CBX6 knockdown. “∗” indicated p < 0.05.

Article Snippet: A pCMV6-entry plasmid containing Myc-DDK (same as Flag)-tagged human CBX6 cDNA (Cat#: RC204166) and a CBX6 Human shRNA Plasmid Kit (Cat#: TF314170) were purchased from OriGene.

Techniques: Over Expression, MTT Assay, Stable Transfection, Transfection, Control, Plasmid Preparation, shRNA, Knockdown

Journal: Molecular Therapy Oncology

Article Title: CBX6 and CA9 as predictive indicators and therapeutic targets in GBM

doi: 10.1016/j.omton.2026.201159

Figure Lengend Snippet: Effects of CBX6 modulation on invasion and migration of U-251 MG cells (A and B) Representative images and cell quantification from transwell migration assays of U-251 MG cells with modulated CBX6 expression compared to corresponding controls. (C and D) Representative images and cell quantification from transwell invasion assays of U-251 MG cells with modulated CBX6 expression compared to respective controls. VC, vector control; OE, CBX6 overexpression; KD, CBX6 knockdown. “∗” Indicates p < 0.05.

Article Snippet: A pCMV6-entry plasmid containing Myc-DDK (same as Flag)-tagged human CBX6 cDNA (Cat#: RC204166) and a CBX6 Human shRNA Plasmid Kit (Cat#: TF314170) were purchased from OriGene.

Techniques: Migration, Expressing, Plasmid Preparation, Control, Over Expression, Knockdown

Journal: Molecular Therapy Oncology

Article Title: CBX6 and CA9 as predictive indicators and therapeutic targets in GBM

doi: 10.1016/j.omton.2026.201159

Figure Lengend Snippet: Effect of human CBX6 overexpression on glioma tumor growth (A and B) Bioluminescent imaging of NSG mice (A) and quantification of bioluminescence signal intensity as fold change (B) on days 4 and 8 ( n = 5). (C) Bioluminescent imaging at week 2 showing reduced Kluc tumor size in mice with human CBX6 overexpression compared to controls ( n = 10). (D and E) Kaplan-Meier survival analysis demonstrating that overexpression of human CBX6 in U-251 MG cells (D) ( n = 5) and Kluc cells (E) ( n = 10) improved mouse survival. (F) Histological analysis of tumor invasion and microsatellite metastasis in CBX6-overexpressing tumors. “∗” Indicates p < 0.05.

Article Snippet: A pCMV6-entry plasmid containing Myc-DDK (same as Flag)-tagged human CBX6 cDNA (Cat#: RC204166) and a CBX6 Human shRNA Plasmid Kit (Cat#: TF314170) were purchased from OriGene.

Techniques: Over Expression, Imaging

Journal: Molecular Therapy Oncology

Article Title: CBX6 and CA9 as predictive indicators and therapeutic targets in GBM

doi: 10.1016/j.omton.2026.201159

Figure Lengend Snippet: CBX6 binds to the CA9 promoter (A) Genes related to tumor cell invasion, proliferation, or migration were selected based on RNA sequencing data from U-251 MG cells with CBX6 knockdown compared to negative controls. (B) qRT-PCR results show that CA9 expression is affected by CBX6 dysregulation in U-251 MG cells, with overexpression or shRNA-mediated knockdown of CBX6, using GAPDH as a reference gene. (C) Gene expression correlation analysis from CGGA reveals an inverse relationship between CBX6 and CA9 expression patterns ( http://www.cgga.org.cn ). (D) Western blot and qRT-PCR data demonstrate changes in CBX6 and CA9 expression in U-251 MG and PBT030 cells under normoxic (N) and hypoxic (H) conditions for 24 and 48 h, using 28S as a reference gene. (E) ChIP assay results from CBX6-overexpressing U-251 MG cells show detection of the CA9 promoter sequence using two primer sets in CBX6/Flag pull-down products compared to a negative IgG control via qRT-PCR. VC, vector control; OE, CBX6 overexpression; KD, CBX6 knockdown. “∗” Indicates p < 0.05.

Article Snippet: A pCMV6-entry plasmid containing Myc-DDK (same as Flag)-tagged human CBX6 cDNA (Cat#: RC204166) and a CBX6 Human shRNA Plasmid Kit (Cat#: TF314170) were purchased from OriGene.

Techniques: Migration, RNA Sequencing, Knockdown, Quantitative RT-PCR, Expressing, Over Expression, shRNA, Gene Expression, Western Blot, Sequencing, Control, Plasmid Preparation

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: SIRT1 deficiency promotes senescence in trophoblast Cells (A) Western blot validates shSIRT1 interference efficiency in HTR8/SVneo cells (B) Western blot detection of the protein levels of p16 and p21; n = 3. (C) SA-β-gal staining of HTR8/SVneo cells; Scale bars: 200 μm; n = 3. (D–F) Levels of (D) IL-6, (E) IL-1αand (F) IL-1β concentration in the cells were measured using the corresponding detection kits; n = 3 independent experiments. Two-tailed t-test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Western Blot, Staining, Concentration Assay, Two Tailed Test

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: SIRT1 alleviates PE-like symptoms in the mouse model (A) Schematic illustration of the experimental design. (B) Representative images of the fetuses of the Control, L-NAME, and L-NAME + SRT2104 groups; Scale bars:1 cm. (C) Systolic blood pressure of pregnant mice in control (n = 6 dam), L-NAME (n = 7 dams), and L-NAME + SRT2104 group (n = 7 dams). (D–F) Fetal weight, (E) crown-rump length,and (F) placental weight at E18.5 in different groups; n = 85 fetuses from 6 dams in the control group, n = 98 fetuses from 7 dams in the L-NAME group, and n = 81 fetuses from 7 dams in the L-NAME + SRT2104 group. (G) H&E staining of placental sections at E18.5. The LZ and JZ areas and the Lz/Jz ratio were quantified; Scale bars: 1000 μm (upper panel), 200um (lower panel); n = 3. (H) H&E staining of maternal kidney sections at E18.5, and measurement of Bowman space; Scale bars: 200 μm; n = 3. LZ, Labyrinth zone; JZ, Junction zone. One-way ANOVA and Tukey’s multiple comparison test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Control, Staining, Comparison

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: SIRT1 rescues placental senescence in the PE mouse model (A) Western blot detection of the protein levels of p16, p21 and SIRT1 in E18.5 mouse placentas from different groups; n = 3. (B) SA-β-gal staining in E18.5 mouse placentas from different groups. Scale bars: 1,000 μm, n = 3 (C,D) Representative immunofluorescence staining of (C) SIRT1(red) and p16 (green) or (D) SIRT1 (red) and p21 (green), in E18.5 placentas from different groups; Nuclei were counterstained with DAPI (blue). Scale bars: 1,000 μm; n = 3. (E–G) Levels of (E) IL-6, (F) IL-1α and (G) IL-1β concentration in the E18.5 mouse placentas from different groups were measured using the corresponding detection kits; n = 3, one-way ANOVA and Tukey’s multiple comparison test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Western Blot, Staining, Immunofluorescence, Concentration Assay, Comparison

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: Elimination of senescent cells alleviates SIRT1 deficiency–induced dysfunction and SASP (A–E) HTR-8/SVneo cells were transfected with shSIRT1 and subsequently treated with PCC1 (5 μM) for 24 h (A) SA-β-gal staining cells; Scale bars: 200 μm; n = 3. (B) Western blot analysis of the protein levels of p16, p21 and SIRT1 in cells; n = 3. (C–E) Levels of (C) IL-6 (D) IL-1α and (E) IL-1β concentration in the cells were measured using corresponding detection kits; n = 3. (F) HTR-8/SVneo cells were transfected with shSIRT1 and then treated with PCC1 (5 μM) or SRT1720 (0.4 uM) for various time periods. Cell viability was measured by CCK8; n = 6. (G–I) HTR-8/SVneo cells were transfected with shSIRT1 and subsequently treated with PCC1 (5 μM) or SRT1720 (0.4 uM) for 24 h. (G) EdU staining; Scale bars: 400 μm; n = 3 (H) Matrigel Transwell assay; Scale bars: 200 μm; n = 3. (I) Wound-healing assay; Scale bars: 400 μm; n = 3. One-way ANOVA and Tukey’s multiple comparison test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Transfection, Staining, Western Blot, Concentration Assay, Transwell Assay, Wound Healing Assay, Comparison

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: Iron metabolism dysregulation correlates with SIRT1 deficiency (A) Volcano plot of the significant differences in gene expression levels between groups. Genes showing the greatest differences were analyzed by (B) GO and (C) KEGG analysis. (D) Heatmap depicting the expression levels of representative genes associated with “intracellular iron ion homeostasis”. (E) Western blot detection of the protein levels of FTH, GPX4, SLC7A11, FPN1 and TFR; n = =3. (F–H) Measurement of intracellular (F) iron concentration, (G) GSH concentration, and (H) MDA concentration using corresponding detection kits; n = 3. (I) Measuring cellular lipid peroxidation by fluorescence microscopy using the C11 BODIPY 581/591 fluorescent probe. Total C11 BODIPY 581/591 (red), oxidized C11 BODIPY 581/591 (green), DAPI (blue) stained nucleus; Scale bar: 100 μm; n = 3. (J) Intracellular Fe 2+ was measured by fluorescence microscopy using the FerroOrange fluorescent probe. FerroOrange (red) stains Fe 2+ and DAPI (blue) stains the nucleus; Scale bar: 200 μm; n = 3. (K) Measuring mitochondrial membrane potential using the JC-1 fluorescent probe. JC-1 aggregates (red), JC-1 monomers (green) and DAPI (blue) stained nucleus; Scale bar: 100 μm; n = 3. (L) Flow cytometry assay for measuring ROS by staining with DCFH-DA, normalized by the number of cells uploaded; n = 3 (M) Transmission electron microscopy images of organelles. Scale bars: 1 µm (left panel) and 500 nm (right panel); N, nucleus; Mi, mitochondria. N = 3, two-tailed t-test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Gene Expression, Expressing, Western Blot, Concentration Assay, Fluorescence, Microscopy, Staining, Membrane, Flow Cytometry, Transmission Assay, Electron Microscopy, Two Tailed Test

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: SIRT1 regulates iron metabolism and senescence in trophoblast cells via p53 deacetylation (A) Reciprocal Co-IP of SIRT1 and p53 in HTR8/SVneo cells. (B) Western blot analysis of ac-p53, p53, and SIRT1; n = 3. (C) Western blot validates si-p53 interference efficiency. (D) Western blot detection of the protein levels of ac-p53, p53, SLC7A11, FPN1, TFR, SIRT1, p16, FTH, p21 and GPX4; n = 3. (E) After treatment, fluorescence images of total C11 BODIPY 581/591 (red) and oxidized C11 BODIPY 581/591 (green) in HTR-8/SVneo cells as indicated; Scale bar: 100 μm; n = 3. (F) After treatment, fluorescence images of Fe 2+ (red) in HTR-8/SVneo cells as indicated; Scale bar: 200 μm; n = 3. (G–I) Levels of (G) IL-6, (H) IL-1α and (I) IL-1β concentration in cells were measured using corresponding detection kits; n = 3. One-way ANOVA and Tukey’s multiple comparison test. All data are presented as the means ± SEM.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques: Co-Immunoprecipitation Assay, Western Blot, Fluorescence, Concentration Assay, Comparison

Journal: Frontiers in Aging

Article Title: The SIRT1–p53 axis drives a ferro-aging-like program and aggravates trophoblast dysfunction in preeclampsia

doi: 10.3389/fragi.2026.1838730

Figure Lengend Snippet: Schematic illustration of the mechanism by which SIRT1 drives a positive feedback loop between iron metabolism and senescence in trophoblasts.

Article Snippet: Lentiviral vectors carrying short hairpin RNA (shRNA) targeting human SIRT1 (GV248) and a negative control shRNA (sh-NC) were purchased from GeneChem (Shanghai, China).

Techniques:

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: IRF7 and pIRF7 are increased in human CLAD (BOS). (A) Representative Western blot images showing IRF7 and pIRF7 in CLAD (BOS) and Stable LTx whole lung lysates. Densitometric analyses of the above blots (fold change relative to Stable LTx) showing protein expression of (B) IRF7 and (C) pIRF7 in CLAD (BOS) and Stable LTx whole lung tissue (Stable LTx n = 3, CLAD (BOS) n = 4). Error bars represent mean ± SEM. ∗∗p < 0.01, unpaired t-test. (D) Representative immunofluorescence images showing H&E, Trichrome, DAPI (blue), IRF7 (magenta), and merged DAPI/IRF7 channels in Stable LTx and CLAD (BOS) lung tissue sections, demonstrating airway-centric distribution and increased IRF7 expression in CLAD (BOS). (E) Corresponding Integrated Intensity/cell of IRF7 in human CLAD (BOS) and Stable LTx specimens. Each data point represents the mean integrated IRF7 intensity/cell averaged across 3–4 image fields per patient (Stable LTx n = 3, CLAD (BOS) n = 4); group data presented as mean ± SEM; ∗p = 0.017, unpaired t-test.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Western Blot, Expressing, Immunofluorescence

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: Virus exposure induces IRF7 and airway fibrogenesis in the air-liquid-interface PBEC model, and IRF7 silencing attenuates it. (A) Representative Western blots showing protein expression of IRF7, α-SMA, and SMAD2/3 in stably expressing control (Ctr) shRNA and IRF7 shRNA primary bronchial epithelial cell (PBEC) cell line air-liquid-interface (ALI) cultures in the presence and absence of Influenza A virus (IAV). Densitometric analyses of the above blots showing protein expression of (B) IRF7, (C) α-SMA, and (D) SMAD2/3 in Ctr shRNA and IRF7 shRNA PBEC cell line ALI cultures (n = 6 each). Error bars represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, Mann–Whitney test for 2 samples, one way Anova with Tukeys multiple comparison test for >3 groups (E) Sircol assay showing fold change soluble collagen content in the ALI culture supernatants from the above experiments (n = 3 each). Error bars represent mean ± SEM. ∗∗p < 0.01, Mann–Whitney test. (F) Representative immunofluorescence images showing DAPI (blue) and MUC5AC (red) of stably expressing control shRNA and IRF7 shRNA cell line ALI cultures exposed to IAV, demonstrating attenuation of airway mucogenesis with IRF7 silencing.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Virus, Western Blot, Expressing, Stable Transfection, Control, shRNA, MANN-WHITNEY, Comparison, Immunofluorescence

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: Virus exposure induces fibrogenesis and IRF7 blockade attenuates it in the human precision-cut lung slice ex vivo model. (A) Representative immunofluorescence images showing IRF7 (red), α-SMA (green), DAPI (blue), and merged channels in a human precision-cut lung slice (PCLS) model in non-treated (NT) and Influenza A virus (IAV)-exposed conditions. (B) Corresponding fold change fluorescence intensity of IRF7 and α-SMA in the above experiment (n = 5–6 each). Error bars represent mean ± SEM. ∗∗p < 0.01, Mann–Whitney test. (C) Representative immunofluorescence images showing IRF7 (red), DAPI (blue), and merged channels, alongside trichrome staining showing collagen deposition, in IAV-exposed stably expressing control shRNA and IRF7 shRNA PCLS slices. (D) % Area of slices positive for trichrome stain in IAV-exposed stably expressing control shRNA and IRF7 shRNA PCLS slices (n = 5–6 each). Error bars represent mean ± SEM. ∗p < 0.05, Mann–Whitney test.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Virus, Ex Vivo, Immunofluorescence, Fluorescence, MANN-WHITNEY, Staining, Stable Transfection, Expressing, Control, shRNA

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: Virus induces IL-33 via IRF7 and IL-33 blockade attenuates virus-induced fibrogenesis. (A) Representative Western blots showing protein expression of IRF7 and IL-33 in stably expressing control shRNA and IRF7 shRNA primary bronchial epithelial cell (PBEC) cell line air-liquid-interface (ALI) cultures in the presence and absence of Influenza A virus (IAV). (B) Densitometry fold change protein expression of IL-33 in control shRNA and IRF7 shRNA PBEC cell line ALI cultures in the presence and absence of IAV (n = 4 each). Error bars represent mean ± SEM. ∗p < 0.05, 1-way ANOVA with Tukey’s multiple comparison post-test. (C) Representative Western blots showing protein expression of IRF7 and α-SMA in stably expressing control shRNA and IRF7 shRNA PBEC cell line ALI cultures in the presence and absence of IAV and IL-33 blockade. (D) Densitometry fold change of α-SMA protein expression in stably expressing control shRNA and IRF7 shRNA PBEC cell line ALI cultures in the presence and absence of IAV and IL-33 blockade (n = 4 each). Error bars represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ns = not significant, 1-way ANOVA with Tukey’s multiple comparison post-test.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Virus, Western Blot, Expressing, Stable Transfection, Control, shRNA, Comparison

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: MMP-9 blockade attenuates virus-mediated fibrogenesis. (A) Fold change mRNA expression of MMP-9 in stably expressing control shRNA and IRF7 shRNA primary bronchial epithelial cell (PBEC) cell line air-liquid-interface (ALI) cultures in the presence and absence of Influenza A virus (IAV) (n = 3 each). Error bars represent mean ± SEM. ∗∗p < 0.01, Mann–Whitney test. (B) MMP-9 (ng/ml of supernatant) concentrations by ELISA in the lower compartment of control shRNA and IRF7 shRNA PBEC cell line ALI culture medium in the presence and absence of IAV (n = 4 each). Error bars represent mean ± SEM. ∗∗∗∗p < 0.0001, 1-way ANOVA with Tukey’s multiple comparison post-test. (C) MMP-9 (ng/ml of supernatant) concentrations by ELISA in the lower compartment of IAV-exposed PBEC cell line ALI model with and without IL-33 blockade (n = 4 each). Error bars represent mean ± SEM. ∗∗∗∗p < 0.0001, ∗∗p < 0.01, ∗p < 0.05, 1-way ANOVA with Tukey’s multiple comparison post-test. (D) MMP-9 (ng/ml of supernatant) concentrations by ELISA in the lower compartment of IAV-exposed stably expressing IRF7 shRNA PBEC ALI model with and without IL-33 blockade (n = 4 each). Error bars represent mean ± SEM. ∗p < 0.05, ns = not significant, 1-way ANOVA with Tukey’s multiple comparison post-test. (E) Representative Western blots showing protein expression of α-SMA in virus-exposed ALI PBEC model with and without MMP-9 blockade. (F) Densitometry fold change protein expression of α-SMA in IAV-exposed ALI PBEC model with and without MMP-9 blockade (n = 5 each). ∗∗∗∗p < 0.0001, ∗∗p < 0.01, ns = not significant, 1-way ANOVA with Tukey’s multiple comparison post-test.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Virus, Expressing, Stable Transfection, Control, shRNA, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Comparison, Western Blot

Journal: eBioMedicine

Article Title: Mechanistic analysis of an IRF7-dependent pathway in virus-induced fibrosis in chronic lung allograft dysfunction

doi: 10.1016/j.ebiom.2026.106285

Figure Lengend Snippet: Schematic representation of the pathway involved in virus-induced airway fibrogenesis. (1 and 2) Influenza virus activates IRF7 via the RIG-1/MAVS/TBK1 axis, and activated IRF7 enters the nucleus and induces expression of IL-33. (3 and 4) IL-33 is secreted from the epithelial cells into the interstitial matrix, where it acts upon neighbouring epithelial cells, resident fibroblasts, and other cell types (paracrine), as well as the same cell (autocrine manner). (5) IL-33 induces MMP-9 expression, and MMP-9 is secreted into the interstitial matrix. (6) MMP-9 cleaves latent TGF-β to its active form. (7 and 8) Activated TGF-β induces SMAD2/3 signalling, driving expression of collagens and α-SMA and culminating in fibrogenesis and extracellular matrix remodelling.

Article Snippet: IRF7 shRNA plasmid complete kit (Origene, cat. TR315487) was used to suppress IRF7 expression in PCLS and were transfected with either IRF7-specific or control shRNA plasmid in the absence of antibiotics in the culture media.

Techniques: Virus, Expressing