vimentin  (Boster Bio)

Journal: Molecular Therapy. Nucleic Acids

Article Title: Nitric oxide-dependent stabilization of vimentin confers chemoresistance in ovarian cancer

doi: 10.1016/j.omtn.2026.102924

Figure Lengend Snippet: iNOS increases sensitivity to cisplatin in ovarian cancer (A) The cell viability of OVCAR8 cells was assessed using RealTime-Glo MT Cell Viability Assay after culturing in increasing concentrations of cisplatin for 72 h. (B) OVCAR8 cells incubated with or without L-NMMA for 48 h, measured by RealTime-Glo MT Cell Viability Assay. (C) Cell viability after either no treatment (control), cisplatin, and a combination of cisplatin and L-NMMA by RealTime-Glo MT Cell Viability Assay. (D) Western blot analysis of vimentin protein expression in ovarian cancer cell lines. (E) Ovarian cancer cell lines were analyzed for VIM mRNA levels by qPCR. (F) OVCAR8 cells were treated with or without L-NMMA (4, 6, 8, and 10 mM) for 48 h, and western blot was used to analyze the effect of L-NMMA on the vimentin protein expression. B-actin was used as a loading control. Band densities were quantified using ImageJ analysis. Error bars, SEM. ∗ p < 0.05; ∗∗ p < 0.01 (compared with the control group, using two-way ANOVA). All experiments were independently repeated three times.

Article Snippet: The following primers for TaqMan Gene Expression Assay were purchased from Thermo Fisher Scientific; human VIM (Cat. #Hs00958111_m1) and human NOS2 (Cat. #Hs01075529_m1).

Techniques: Viability Assay, Incubation, Control, Western Blot, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Nitric oxide-dependent stabilization of vimentin confers chemoresistance in ovarian cancer

doi: 10.1016/j.omtn.2026.102924

Figure Lengend Snippet: iNOS knockout increased chemosensitivity and impaired cell motility and migration of ovarian cancer cells (A) Immunoblotting showing reduced iNOS (left) and vimentin (right) expression levels following NOS2 knockdown in the OVCAR8 cells. (B) NOS2 knockdown sensitized OVCAR8 cells to cisplatin, reducing its IC 50 value. Log-logistic model was used to analyze the data. The group comparison between control group and KO-1 had a p value = 8.98e-07. The comparison between control and KO-2 had a p value = 0.0132. The detected EC50 for control group was 0.8554, for KO-1 was 0.6037, for KO2 was 0.7688. (C and D) Analysis of reduced protein (left) and mRNA (right) expression of iNOS and vimentin following siRNA transfection in OVCAR8 and A2780cis cells for 72 h, assessed by western blot and RT-qPCR. (E) OVCAR8 cells were transfected with 2 different siRNAs or the scramble siRNA and were assessed for migration using the scratch wound assay. The area of the wound was measured at 0, 12, 24, and 36 h by the IncuCyte live-cell analysis system. Two-way repeated measures ANOVA was used to analyze the data. After 6 h, siRNA1 had p value = 1, siRNA2 had p value = 0.23. After 12 h, siRNA1 had p value = 0.71 and siRNA 2 had p value = 0.16. After 24 h, siRNA1 had a p value = 0.39 and siRNA2 had a p value = 0.05. After 36 h, siRNA1 had a p value = 0.86 and siRNA2 had a p value = 0.04. (F) NOS2 KO-1 and KO-2 OVCAR8 cells formed significantly fewer colonies compared to parental OVCAR8. The experiment was performed in triplicate with three biological replicates. Statistical analysis was conducted with two-way ANOVA. ∗ p < 0.05 and ∗∗ p < 0.01. (G) Silencing of NOS2 by two different siRNAs significantly reduced the number of colonies formed by OVCAR8 cells. Clonogenic growth was measured after 10 days, quantified using ImageJ, and represented as a bar graph (mean ± SEM). The experiment was performed in triplicate with three biological replicates. Statistical analysis was conducted with two-way ANOVA. ∗ p < 0.05 and ∗∗ p < 0.01. All experiments were independently repeated two to three times.

Article Snippet: The following primers for TaqMan Gene Expression Assay were purchased from Thermo Fisher Scientific; human VIM (Cat. #Hs00958111_m1) and human NOS2 (Cat. #Hs01075529_m1).

Techniques: Knock-Out, Migration, Western Blot, Expressing, Knockdown, Comparison, Control, Transfection, Quantitative RT-PCR, Scratch Wound Assay Assay, Cell Analysis

Journal: Molecular Therapy. Nucleic Acids

Article Title: Nitric oxide-dependent stabilization of vimentin confers chemoresistance in ovarian cancer

doi: 10.1016/j.omtn.2026.102924

Figure Lengend Snippet: L-NMMA promotes vimentin destabilization by enhancing its ubiquitination (A) CHX chase assay showing vimentin protein levels in OVCAR8 cells treated with 10 mM L-NMMA at different time points. (B) Vimentin ubiquitination was assessed in OVCAR8 cells treated with L-NMMA for 24 h, in the presence of the proteasome inhibitor MG-132 (10 μM) for the final 4 h, followed by immunoprecipitation and western blot using an anti-ubiquitin antibody. (C) Measurement of vimentin S-nitrosylation levels was performed by immunoprecipitation in OVCAR8 cells. (D) Western blot analysis of vimentin expression in OVCAR8 cells treated with 10 mM L-NMMA alone or in combination with MG132 at 1, 5, or 10 μM. (E) Tumor growth and proliferation were monitored in mice bearing parental and NOS2 KO OVCAR8 tumors, evaluated by ROI measurements every 4 days ( n = 6). (F) Kaplan-Meier survival curves of mice bearing parental and NOS2 KO OVCAR8 tumors following cisplatin treatment ( n = 6). Data are presented as mean ± SEM. Statistical analysis was performed using two-way ANOVA for growth curves and the Kaplan-Meier method for survival analysis ( p < 0.05, ∗ p < 0.01). Experiments were performed in duplicate or triplicate.

Article Snippet: The following primers for TaqMan Gene Expression Assay were purchased from Thermo Fisher Scientific; human VIM (Cat. #Hs00958111_m1) and human NOS2 (Cat. #Hs01075529_m1).

Techniques: Ubiquitin Proteomics, Immunoprecipitation, Western Blot, Expressing

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry

Journal: iScience

Article Title: MFAP5 + synovial fibroblasts drive LOX upregulation to promote osteoarthritis progression

doi: 10.1016/j.isci.2026.116286

Figure Lengend Snippet: MFAP5 upregulation in synovium of end-stage OA patients correlated with aggressive synovitis (A) Representative images of H&E staining of synovial tissues across three groups of humans. Scale bars, 100 μm. (B) Distribution of KSS in synovial tissues across the three human groups ( n = 3–5). (C) The colocalization pixel ratio of MFAP5 and vimentin was quantified in synovial tissues across three groups of humans ( n = 4). (D) Representative immunofluorescence images of MFAP5 and vimentin colocalization in synovial tissues. Scale bars, 500 (top) and 20 μm (bottom). (E) Fluorescent intensity of MFAP5 in synovial tissues across the three human groups ( n = 3–4). (F) The mRNA level of MFAP5 in synovial tissues across the three human groups was measured using real-time qPCR assay ( n = 3–5). (G and H) Correlation between KSS and MFAP5 fluorescent intensity (G)/mRNA level (H) in synovial tissues. (I) Representative immunofluorescence images of FBN1 and vimentin colocalization in synovial tissues. Scale bars, 500 (top) and 20 μm (bottom). (J) The colocalization pixel ratio of FBN1 and vimentin was quantified in synovial tissues across three groups of humans ( n = 4). (K) Fluorescent intensity of FBN1 in synovial tissues across the three human groups ( n = 3–4). (L) The mRNA level of FBN1 in synovial tissues across the three human groups was measured using real-time qPCR assay ( n = 3–5). (M and N) Correlation between KSS and FBN1 fluorescent intensity (K)/mRNA level (L) in synovial tissues. Data were presented as mean ± SEM. Statistical significance was calculated using one-way ANOVA followed by Tukey’s multiple comparisons test (B, C, E, F, and J–L) and Spearman's rank correlation analysis (G, H, M, and N). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies, including vimentin (GB11192, Servicebio, China), FBN1 (860327, Zenbio, China), MFAP5 (15727-1-AP, Proteintech, China), and CD4 (GB150062-50, Servicebio, China).

Techniques: Staining, Immunofluorescence

Journal: iScience

Article Title: MFAP5 + synovial fibroblasts drive LOX upregulation to promote osteoarthritis progression

doi: 10.1016/j.isci.2026.116286

Figure Lengend Snippet: MFAP5 + SFs promoted collagen deposition in synovium (A) Dendrogram showing the hierarchy organization of MEGENA network modules in GSE176308 . The top layer represented the root and largest parent modules, which branched hierarchically into smaller child modules. (B) Density plots displaying c1_9 module scores across SF clusters, with color-coded density values; red indicated higher density, and blue indicated lower density. (C) Network of pathways and genes involved in the c1_9 module, where a two-point line indicated that a gene was part of a specific pathway. (D) Raincloud plot showing c1_9 score in GSE152805 . (E and F) Gene set enrichment analysis showed collagen pathways were upregulated in MFAP5 + SFs in both GSE176308 (E) and GSE152805 (F). (G) Correlation coefficients between the expression of MFAP5 and one ECM pathway score across three datasets. (H) Violin plots of ECM binding (left) and ECM organization (right) scores across SF clusters in GSE176308 . Color-coded by SF clusters. (I) Density plots of ECM binding and ECM organization scores across SF clusters in SCP469 and GSE152805 . (J) Comparison of significant ligand-receptor pairs from SF clusters to T cells in GSE152805 . (K) Immunofluorescence images of MFAP5, CD4, and vimentin colocalization in synovial tissues. Scale bars, 200 or 20 μm. Statistical significance was calculated using two-tailed Wilcoxon rank-sum test (D and H) and Spearman's rank correlation analysis (G). ∗∗∗ p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies, including vimentin (GB11192, Servicebio, China), FBN1 (860327, Zenbio, China), MFAP5 (15727-1-AP, Proteintech, China), and CD4 (GB150062-50, Servicebio, China).

Techniques: Expressing, Binding Assay, Comparison, Immunofluorescence, Two Tailed Test

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 knockdown inhibits gene signatures characteristic of CMS4 colorectal cancer. (A) qRT-PCR analysis was applied to examine the relative mRNA expression of genes related to epithelial–mesenchymal transition (EMT), complement, angiogenesis, and immunosuppression. GAPDH was a normalization control. (B) Protein expression and quantification results of SNAIL, Vimentin, and E-cadherin in colorectal cancer cells were determined by western blotting. (C) Immunofluorescence analysis of the levels of E-cadherin (the epithelial marker, red) and Vimentin (the mesenchymal marker, green) proteins in the control and PLXNC1-silenced LoVo cells. The nuclei were stained with Hoechst (blue). Scale bars = 75 μm ∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05.

Article Snippet: Fixed cells were incubated with primary antibody against E-cadherin (1:50; sc-8426; Santa Cruz Biotechnology, California, USA) or vimentin (1:100; BM0135; Boster, Wuhan, China) at 4 °C overnight, followed by incubation with secondary antibodies (1:200; Boster, Wuhan, China) at room temperature for 1 h. The nuclei of cells were stained with Hoechst for 10 min.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Control, Western Blot, Immunofluorescence, Marker, Staining

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 promotes tumor growth and metastasis in vivo . (A) Magnetic resonance imaging (MRI) assessment of wild-type (WT) mice carrying subcutaneous tumors. (B) Tumor volumes calculated by MRI. The tumor volume = area of tumor in each slice × slice thickness. (C) Representative images of tumors from WT mice after subcutaneous inoculation of SL4 cells infected with the control vector or sh-PLXNC1 vector. (D) Mice were killed 14 days following subcutaneous injection, and tumors were dissected and weighed. (E) Bioluminescence images showing tumor metastasis by tracking luciferase-expressing SL4 cells in sh-NC and sh-PLXNC1 groups. (F) Histogram showing the bioluminescent signal intensity analyzed by the IVIS System. (G) MRI inspection of hepatic tumor metastasis of colon cancer after intrasplenic injection of SL4 cells in sh-NC and sh-PLXNC1 groups. (H) Gross examination of hepatic tumor metastasis of colon cancer after intrasplenic injection of SL4 cells in sh-NC and sh-PLXNC1 groups. (I) Mice were sacrificed on day 10 after intrasplenic injection, and livers were excised and weighed. (J) Immunohistochemical staining was used to detect the expression levels of PCNA, CD31, TGF-β, vimentin, β-catenin, and E-Cadherin in tumor tissues from sh-NC and sh-PLXNC1 groups. Scale bars = 50 μm. (K) Quantification of immunohistochemical staining in subcutaneous tumor tissues from sh-NC and sh-PLXNC1 groups. (L) Immunohistochemical staining showing expression of epithelial–mesenchymal transition (EMT) markers in liver metastasis tumor tissues from sh-NC and sh-PLXNC1 groups. (M) Quantification of immunohistochemical staining in liver metastasis tumor tissues from sh-NC and sh-PLXNC1 groups. ∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05; ns, not significant.

Article Snippet: Fixed cells were incubated with primary antibody against E-cadherin (1:50; sc-8426; Santa Cruz Biotechnology, California, USA) or vimentin (1:100; BM0135; Boster, Wuhan, China) at 4 °C overnight, followed by incubation with secondary antibodies (1:200; Boster, Wuhan, China) at room temperature for 1 h. The nuclei of cells were stained with Hoechst for 10 min.

Techniques: In Vivo, Magnetic Resonance Imaging, Infection, Control, Plasmid Preparation, Injection, Luciferase, Expressing, Immunohistochemical staining, Staining

Journal: Materials Today Bio

Article Title: Extracellular biogenic nanoscale mitochondria reprogram the wound microenvironment via ROS scavenging independent of cellular uptake

doi: 10.1016/j.mtbio.2026.103023

Figure Lengend Snippet: MSC-mt alleviates oxidative stress and promote tissue regeneration during wound healing (A) In vivo imaging showing the spatial–temporal persistence of fluorescently labeled MSC-mt (mtH) at the wound site at indicated time point, indicating transient but sustained early presence after topical application. (B) Measurement of ATP levels in peri-wound tissues on PWD8 showed enhanced local metabolic activity following mtH treatment. n = 5 ∼ 6 per group. (C) Quantification of malondialdehyde (MDA) levels in peri-wound tissues on PWD8 indicated reduced lipid peroxidation and oxidative stress in both MSC-mt–treated wounds. n = 5 ∼ 6 per group. (D) Laser speckle contrast imaging of blood perfusion at the wound site on PWD8 showed improved microvascular perfusion following mtH treatment. n = 5 per group. (E) Representative immunofluorescence images and quantification of CD31 expression in peri-wound tissues on PWD8, indicating enhanced angiogenesis in mtH–treated wounds. n = 6 per group. (F) Quantitative PCR analysis of angiogenesis-related gene expression in peri-wound tissues on PWD8, indicating transcriptional activation of pro-angiogenic programs following mtH treatment. n = 3 ∼ 5 per group. (G-H) Representative immunohistochemical staining and quantification of Col1a1 in wound tissues on PWD8, showing increased collagen synthesis and matrix remodeling in mtH–treated wounds. n = 6 per group. Scale bar = 100 μm. (I-J) Representative immunofluorescence staining and quantification of Vimentin and TUNEL in wound tissues on PWD8, indicating reduced fibroblast apoptosis following mtH treatment. n = 6 per group. Scale bar = 20 μm. (K-L) Representative immunofluorescence staining and quantification of Vimentin and 8-hydroxyguanosine (8-OHG) in wound tissues on PWD8, indicating attenuated oxidative DNA damage in fibroblasts following mtH treatment. n = 6 per group. Scale bar = 20 μm. Data are presented as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against CD31 (Servicebio, Cat# GB120005 , 1:200), Vimentin (CST, Cat# 5741, 1:200), and 8-hydroxyguanosine (8-OHG, Rockland, Cat# 200-301-A99, 1:200).

Techniques: In Vivo Imaging, Labeling, Activity Assay, Imaging, Immunofluorescence, Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Activation Assay, Immunohistochemical staining, Staining, TUNEL Assay