mmp9  (R&D Systems)

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: MMP-9 , a factor that promotes Vasculogenic mimicry, is highly expressed in CC and is associated with poor prognosis. (A) CC database of TCGA was used to analyze key factors associated with VM. (B) Association of Sox2 expression with overall survival in CC (log-rank test). (C) Association of MMP-9 expression with overall survival in CC (log-rank test). (D) Panoramic scans after immunohistochemical detection of MMP-9 and H&E staining in samples from cancerous and paracancerous tissues from subjects with CC. Scale bar, 50 µm. Original magnification, ×20. (E) Protein levels of MMP-9 in 20 paired samples, with the MMP-9 level in CC tissue expressed compared with that in the paired normal tissue. (F) Expression levels of MMP-9 mRNA in 44 paired CC and paracancerous tissues, with MMP-9 expression in CC tissue expressed compared with that in the paired normal tissue. (G) Comparison of the average expression levels of MMP-9 mRNA in CC tissues compared with paracancerous tissues. (H) HeLa and SiHa cells were incubated under hypoxia (0.1% O 2 ) and proteins collected at 24, 48 and 72 h for western blotting of ALDH1, EPHA2, MMP-9 and GAPDH. ImageJ was used to semi-quantify western blotting signals from HeLa (I) and SiHa (J) cells. GAPDH served as an internal reference. *P<0.05, **P<0.01 and ***P<0.001. MMP-9, matrix metalloproteinase 9; VM, vasculogenic mimicry; ALDH1, aldehyde dehydrogenase 1; EPHA2, ephrin type-A receptor 2; TCGA, The Cancer Genome Atlas; Sox2, SRY-box transcription factor 2; CC, cervical cancer; CESC, cervical squamous cell carcinoma.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison, Incubation, Western Blot

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 promotes Vasculogenic mimicry, invasion and migration of CC cells under hypoxic conditions. (A) Expression levels of NSUN2 in CC cells lines (HeLa, SiHa, CaSki, C33A and HT-3) and in a normal cervical cell line (HaCaT). (B) RT-qPCR was used to determine relative expression levels of NSUN2 mRNA in HeLa and SiHa cells after transfection of shRNAs and incubation under hypoxia for 24 h. (C) Relative expression levels of MMP-9 mRNA in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 24 h. (D) Dot blot assay analysis of m 5 C expression levels in HeLa and SiHa cells after 48 h of hypoxia culture following transfection with NSUN2 knockdown plasmids. (E) Semi-quantitative analysis of dot blot results in HeLa cells. (F) Semi-quantitative analysis of dot blot results in SiHa cells. (G) Western blotting was used to investigate NSUN2 and MMP-9 protein levels in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 48 h. ImageJ was used to semi-quantify western blotting bands for (H) NSUN2 and (I) MMP-9 protein levels. (J) 2D tube-forming assays of HeLa and SiHa cells transfected with an NSUN2 -interfering plasmid and incubated under hypoxia. Scale bar, 100 µm. Original magnification, ×4. (K) Assay displayed in panel (J) was quantified using Image Pro. (L) Invasion assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (M) Assay displayed in panel was quantified using Image Pro. (N) Migration assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (O) Assay displayed in panel (N) was quantified using Image Pro. *P<0.05, **P<0.01 and ***P<0.001. VM, Vasculogenic mimicry; CC, cervical cancer; IHC, immunohistochemistry; NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; RT-qPCR, reverse transcription-quantitative PCR; IHC, immunohistochemistry; CaSki, human cervical cancer cell line with intestinal metastasis; C33A, human cervical cancer cell line; HaCaT, human skin keratinocytes cell line; HeLa, human cervical cancer cell line; HT-3, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, Incubation, Plasmid Preparation, Dot Blot, Knockdown, Quantitative Dot Blot, Western Blot, Invasion Assay, Control, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cloning, shRNA, Negative Control

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 increases the stability of MMP-9 mRNA. (A) A positive correlation was observed between NSUN2 and MMP-9 mRNA expression levels in 44 pairs of samples from subjects with CC. Enrichment of the m 5 C modification of MMP-9 mRNA in HeLa (B) and SiHa (C) Cells were measured with anti-m 5 C methylated-RNA IP assays. Interaction of NSUN2 with MMP-9 mRNA in (D) HeLa and (E) SiHa cells was measured with anti- NSUN2 RNA IP assays. Stability of MMP-9 mRNA after interference with and overexpression of NSUN2 was measured in (F) HeLa and (G) SiHa cells. (H) A model illustrating the proposed mechanism by which NSUN2-mediated stabilization of MMP-9 mRNA promotes Vasculogenic mimicry in CC. NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; HeLa, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; IP, immunoprecipitation; CC, cervical cancer; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Modification, Methylation, Over Expression, Plasmid Preparation, Cloning, shRNA, Immunoprecipitation, Negative Control

Journal: bioRxiv

Article Title: Characterization of a chronic UV-induced photoaging mouse model: insights into skin barrier dysfunction, extracellular matrix remodeling, and altered adipogenesis

doi: 10.64898/2026.04.10.712660

Figure Lengend Snippet: (A) TEWL and (B) hydration of the SC of SKH-1 mice were recorded once a week during the entire experiment via the MultiProbe Adapter System MPA 580 (Courage + Khazaka electronic GmbH, Germany). (C) Collagen content in the irradiated and nonirradiated mice was quantified at weeks 2, 4, 6, and 8 via histological analysis. (D) MMP9 expression was analyzed in the skin of nonirradiated and irradiated mice after 8 weeks of irradiation. (E) Hyaluronic acid (HA) levels were determined at weeks 2, 4, 6, and 8 via HPLC-MS. (F-K) Gene expression analysis of selected HA-related markers was performed, (L-N) the gene expression of leptin, adiponectin, and cathelicidin was evaluated, (O) adipocyte size distribution was assessed via histological analysis in the skin of nonirradiated and irradiated mice after 8 weeks of irradiation. Data represent mean values ± standard deviation ( n = 5-7 ) ( * p < 0.05, ** p < 0.001, *** p < 0.0001 compared to nonirradiated control).

Article Snippet: Gene expression was measured via qPCR via the following TaqMan gene expression assays (Thermo Fisher Scientific): IL-1β Mm00434228_m1, MMP9 Mm00442991_m1, COL1A1 Mm00801666_g1, PPARγ Mm00440940_m1, adiponectin Mm04933656_m1, leptin Mm00434759, HAS2 Mm00515089_m1, HAS3 Mm00515092_m1, CEMIP Mm00472921_m1, CEMIP2 Mm00459599_m1, HYAL1 Mm00480053_m1, HYAL2 Mm00477731_m1, CD44 Mm01277161_m1, HMMR Mm00469183_m1, TNFAIP6 Mm00493736_m1 and Ribosomal Protein L13a - RPL13A Mm01612986_g1 and TaqMan Fast Advanced Master Mix (Life Technologies), in duplicate, via a Quant Studio 3 qPCR instrument (Thermo Fisher Scientific).

Techniques: Irradiation, Expressing, Gene Expression, Standard Deviation, Control

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: Downregulation of circ_0103896 is associated with the phenotypic switch of hBVSMCs from a contractile to a synthetic state. ( A ) Expression of circ_0103896 in hBVSMCs following stimulation with PDGF-BB (10 ng/mL) for the indicated times, determined by RT-qPCR. ( B ) hBVSMCs were transfected with siRNA or overexpression plasmids targeting circ_0103896, or corresponding controls, and circ_0103896 expression was quantified by RT-qPCR. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. ( C-E ) Representative bar graphs ( C, E ) and immunoblots ( D ) showing expression of α-SMA and SM22α at the transcriptional ( C ) and translational ( D, E ) levels, assessed by RT-qPCR ( C ) and western blotting ( D, E ), respectively, in differentially treated hBVSMCs. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. ( F-I ) Bar graphs showing the expression of inflammatory cytokines TNF-α ( F ), IL1B ( G ), IL6 ( H ), and iNOS ( I ), in differentially treated hBVSMCs. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. ( J, K ) Representative immunoblots ( J ) and quantification ( K ) showing MMP2 and MMP9 protein levels in differentially treated hBVSMCs. ( L ) Cell proliferation of hBVSMCs under the indicated conditions was assessed using the MTT assay. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. ( M, N ) Representative images ( M ) and quantification ( N ) showing migration of hBVSMCs with or without circ_0103896 silencing or overexpression following PDGF-BB treatment. ∗∗∗∗ P < 0.0001. ( O, P ) Representative histograms ( O ) and quantification ( P ) showing apoptosis of hBVSMCs with or without circ_0103896 silencing or overexpression following PDGF-BB treatment. ∗∗ P < 0.01. Data are presented as mean ± SEM from ≥3 independent biological replicates (separate hBVSMCs cultures). Technical replicates (triplicate wells/qPCR reactions) were averaged within each experiment. Comparisons between two groups were analyzed using an unpaired Student's t -test; multiple-group comparisons were analyzed by one-way ANOVA followed by Tukey's post hoc test. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Over Expression, Western Blot, MTT Assay, Migration

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: circ_0103896 expression is negatively correlated with IA severity in mice. ( A ) Representative H&E-stained images showing vessel wall integrity in IA mice subjected to different treatments. ( B ) Quantification of IA severity based on aneurysmal scores in mice indicated treatments. ∗∗ P < 0.01 and ∗∗∗∗ P < 0.0001. ( C ) Grade distribution of aneurysms in IA mice with differential treatments, displayed as pie charts. ( D-G ) Expression levels of inflammatory mediators, including TNF-α ( D ), IL1B ( E ), MMP9 ( F ), and MMP2 ( G ), in cerebral arteries of IA mice under the indicated treatments. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. In vivo data are represented as mean ± SEM, with each mouse representing one biological replicate ( n = 6 per group unless otherwise indicated). Aneurysmal scores were analyzed using one-way ANOVA followed by Tukey's post hoc test; The IA grade distributions were analyzed using the chi-square test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Mice were randomly allocated to groups using a computer-generated sequence stratified by baseline body weight. Histological scoring and grade assignment were performed on coded samples by two independent investigators blinded to treatment groups. Sample sizes ( n = 6 per group) were determined a priori based on a power analysis for the aneurysmal score (α = 0.05, power = 0.8, expected mean difference = 1.3, SD = 0.8).

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: Expressing, Staining, In Vivo, Generated, Sequencing

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: Suppression of circ_0103896 promotes the phenotypic switch of hBVSMCs from a contractile to a synthetic state via miR-432-5p. hBVSMCs were transfected with control vector (p-NC), circ_0103896 overexpression vector (p-Circ), p-Circ and miR-432–5p mimic (p-Circ + mimic), siRNA negative control (si-NC), siRNA against circ_0103896 (si-Circ), and siRNA against circ_0103896 combined with anti-miR-432–5p (si-Circ + Anti-miR). Following transfection, cells were treated with PDGF-BB for 24 h. ( A ) RT-qPCR analysis of miR-432-50 expression after differential transfection. ∗∗∗ P < 0.001. ( B, C ) Transcriptional levels of α-SMA ( ACTA2 ) ( B ) and SM-22α ( TAGLN ) ( C ) in hBVSMCs after indicated treatments. ∗ P < 0.05 and ∗∗∗ P < 0.001. ( D, E ) Representative western blots ( D ) and quantification ( E ) of α-SMA and SM22α protein expression. ∗ P < 0.05 and ∗∗ P < 0.01. ( F, G ) mRNA expression of inflammation-associated cytokines: TNF-α and IL1B ( F ) and IL6 and iNOS ( G ) in hBVSMCs under indicated treatments measured by RT-qPCR. ∗∗∗ P < 0.001. ( H, I ) Representative western blots ( H ) quantification ( I ) of MMP2 and MMP9 protein levels in hBVSMCs after indicated treatments. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. ( J ) Cell proliferation determined with or without indicated transfection under PDGF-BB treatment. ∗∗∗ P < 0.001. ( K, L ) Representative images ( K ) and quantification ( L ) of hBVSMC migration. ∗∗∗ P < 0.001. ( M, N ) Flow cytometry analysis of apoptosis in hBVSMCs under differential treatments. ∗∗∗ P < 0.001. Data are presented as mean ± SEM from ≥3 independent biological experiments. Technical replicates (triplicate wells or qPCR reactions) were averaged within each experiment. Statistical significance was assessed using one-way ANOVA with Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: Transfection, Control, Plasmid Preparation, Over Expression, Negative Control, Quantitative RT-PCR, Expressing, Western Blot, Migration, Flow Cytometry

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: FTO is regulated by the circ_0103896/miR-432–5p axis in hBVSMCs. ( A ) Schematic highlighting the predicted binding sites of miR-432–5p within the 3′UTR of FTO. ( B ) Luciferase essay in hBVSMCs comparing WT and mutant FTO 3′UTRs following miR-432–5p mimic transfection. ∗∗∗ P < 0.001. ( C-E ) FTO expression in hBVSMCs after transfection with circ_0103896 overexpression vectors or miR-432–5p mimics, assessed by RT-qPCR ( C ) and Western blot ( D, E ). ∗∗∗ P < 0.001. ( F-I ) Protein expression of α-SMA ( F, G ), SM22α ( F, G ), and MMP2 and MMP9 ( H, I ), in hBVSMCs transfected with circ_0103896 or miR-432–5p mimics. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. ( J-N ) Assessment of hBVSMC proliferation ( J ), migration ( K, L ), and apoptosis ( M, N ) after transfection with FTO plasmids or miR-432–5p mimics, using MTT assay, Transwell migration, and cell apoptosis analysis. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. Data are represented as mean ± SEM from ≥3 independent biological replicates. qPCR and luciferase assays were performed in technical triplicate. Statistical comparisons were conducted using an unpaired Student's t -test for two-group comparisons or one-way ANOVA with Tukey's post hoc test for multiple groups. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: Binding Assay, Luciferase, Mutagenesis, Transfection, Expressing, Over Expression, Quantitative RT-PCR, Western Blot, Migration, MTT Assay

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: FTO mediates m 6 A modification of circ_0103896, establishing a positive feedback loop that reinforces circ_0103896 expression. ( A, B ) Global RNA m 6 A levels in hBVSMCs after FTO overexpression or silencing, measured by MeRIP-qPCR ( A ) and m 6 A dot-blot assay ( B ). Dot-blot signals were normalized to methylene blue staining. ∗∗ P < 0.01. ( C ) MeRIP analysis of circ_0103896 m 6 A modification in hBVSMCs following FTO overexpression or silencing. ∗∗ P < 0.01. ( D ) RT-qPCR measurement of circ_0103896 expression in hBVSMCs after FTO overexpression or silencing. ∗ P < 0.05 and ∗∗∗ P < 0.001. ( E ) RIP analysis showing binding between FTO and circ_0103896 in hBVSMCs. ∗∗∗ P < 0.001. ( F, G ) Representative western blots ( F ) and quantification ( G ) blots of α-SMA and SM22α protein levels after indicated transfections. ∗∗∗ P < 0.001. ( H, I ) Representative western blots ( H ) and quantification ( I ) of MMP2 and MMP9 protein levels in hBVSMCs after indicated transfections. ∗∗∗ P < 0.001. ( J ) MTT assay assessing hBVSMC proliferation following circ_0103896 silencing, FTO overexpression, or combined treatments. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. ( K, L ) Representative images ( K ) and quantification ( L ) of hBVSMC migration after circ_0103896 silencing, FTO overexpression, or combined treatments. ∗∗∗ P < 0.001. ( M, N ) Representative histograms ( M ) and quantification ( N ) of apoptosis in hBVSMCs after circ_0103896 silencing, FTO overexpression, or combined treatments. ∗∗ P < 0.01 and ∗∗∗ P < 0.001. Data are presented as mean ± SEM from ≥3 independent biological replicates. MeRIP and dot-blot assays were performed in technical triplicate for each sample. Comparisons were analyzed using one-way ANOVA with Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: Modification, Expressing, Over Expression, Dot Blot, Staining, Quantitative RT-PCR, Binding Assay, Western Blot, Transfection, MTT Assay, Migration

Journal: Non-coding RNA Research

Article Title: circ_0103896/miR-432–5p/FTO feedback loop suppresses the formation and progression of intracranial aneurysm

doi: 10.1016/j.ncrna.2025.11.001

Figure Lengend Snippet: circ_0103896 suppresses IA formation and progression in vivo via the circ_0103896/miR-432–5p/FTO feedback loop. ( A ) Representative H&E-stained images illustrating vessel wall integrity in IA mice under different treatments. ( B ) Representative bar graph depicting the aneurysmal scores of IA mice under differential treatments. ∗∗ P < 0.01 and ∗∗∗∗ P < 0.0001. ( C ) Pie charts depicting the distribution of IA grades across treatment groups. ( D-G ) Representative bar graphs showing mRNA expression of inflammatory mediators, TNF-α ( D ), IL1B ( E ), MMP9 ( F ), and MMP2 ( G ), in cerebral arteries of differentially treated IA mice. ∗∗∗ P < 0.001. Each mouse represents one biological replicate ( n = 6 per group unless otherwise indicated). Data are presented as mean ± SEM. Aneurysmal scores and cytokine expression were compared using one-way ANOVA with Tukey's post hoc test; The IA grade distributions were analyzed by chi-square test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Mice were randomly assigned to groups using a computer-generated sequence stratified by baseline body weight. Histological scoring, grade assignment, and cytokine quantification were performed on coded samples by investigators blinded to group allocation. Group sizes ( n = 6 per group) were determined a priori based on power calculation for the aneurysmal score (α = 0.05, power = 0.8, expected mean difference = 1.3, SD = 0.8).

Article Snippet: All antibodies were purchased from Cell Signaling Technology (USA): FTO (45980S, 1:1000), α-SMA (14968S, 1:1000), SM22α (52011S, 1:1000), MMP2 (14351S, 1:1000), MMP9 (24317S, 1:1000), and GAPDH (2118L, 1:1000).

Techniques: In Vivo, Staining, Expressing, Generated, Sequencing