Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: ACVR1C is a direct target gene of miR-488-3p. (A) Venn analysis of the target genes of miR-488-3p predicted by TargetScan, miRDB and miRWalk. (B) Venn analysis of the potential target genes of miR-488-3p and upregulated mRNAs in LSCC. (C) Quantitative PCR analysis of the mRNA levels of ACVR1C, ABCA12, TNFSF11 and SYT14 in LSCC cells transfected with miR-488-3p or NC mimics. (D) Western blotting of the protein levels of ACVR1C. (E) Schematic of ACVR1C 3′ UTR sequence containing WT and Mut miR-488-3p binding sites. Luciferase reporter assay measured the luciferase activity of ACVR1C 3′ UTR when FD-LSC-1 cells were transfected with miR-488-3p (F) mimics or (G) inhibitor. ** P<0.01 and *** P<0.001. ACVR1C, activin A receptor type 1C; miR, microRNA; LSCC, laryngeal squamous cell carcinoma; NC, negative control; UTR, untranslated region; WT, wild type; Mut, mutant; N.S., not significant.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: Real-time Polymerase Chain Reaction, Transfection, Western Blot, Sequencing, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Negative Control, Mutagenesis

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: ACVR1C knockdown inhibits LSCC cell proliferation, migration, invasion and epithelial-mesenchymal transition. (A) Analysis of ACVR1C expression in RNA-seq data ( GSE127165 ). (B) Quantitative PCR and (C) western blot analysis of the mRNA and protein expression levels of ACVR1C in AMC-HN-8 and FD-LSC-1 cells transfected with si-ACVR1C-1 and 2 or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. (D) Cell Counting Kit-8 and (E) colony formation assays investigated the proliferative ability of LSCC cells transfected with si-ACVR1C or si-NC. In the Transwell assay, ACVR1C knockdown significantly suppressed the (F) migration and (G) invasion of LSCC cells. (H) Western blot analysis of the protein levels of E-cadherin, N-cadherin and vimentin in LSCC cells transfected with si-ACVR1C or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. Scale bar, 100 µ m. * P<0.05, ** P<0.01 and *** P<0.001. ACVR1C, activin A receptor type 1C; LSCC, laryngeal squamous cell carcinoma; NC, negative control; si, small interfering RNA.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: Knockdown, Migration, Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Cell Counting, Transwell Assay, Negative Control, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: miR-488-3p suppresses LSCC cell proliferation, migration and invasion by targeting ACVR1C. (A) Western blot analysis of ACVR1C protein in FD-LSC-1 and AMC-HN-8 cells transfected with OE-ACVR1C or control empty plasmid (OE-NC). Flag antibodies were used to detect the Flag-tagged ACVR1C proteins. (B) Western blot analysis of ACVR1C protein expression in FD-LSC-1 and AMC-HN-8 cells co-transfected with OE-NC and NC mimics (Control), OE-NC and miR-488-3p mimics (miR-488-3p mimics), NC mimics and ACVR1C OE (ACVR1C OE) or miR-488-3p mimics and ACVR1C OE (miR-488-3p mimics + ACVR1C OE). (C) Cell Counting Kit-8 and (D) colony formation assays detected the proliferative ability of LSCC cells in different groups. (E and F) Transwell assays determined the migration and invasion of LSCC cells in different groups. Scale bar, 100 µ m. ** P<0.01 and *** P<0.001. ACVR1C, activin A receptor type 1C; miR, microRNA; LSCC, laryngeal squamous cell carcinoma; OE, overexpression.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: Migration, Western Blot, Transfection, Control, Plasmid Preparation, Expressing, Cell Counting, Over Expression

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: Effect of miR-488-3p on tumor growth in vivo . (A) Image of xenograft tumors after intratumoral injection of miR-488-3p or NC agomir. The xenograft tumor volume was measured to plot the growth curve. (B) Final weight of xenograft tumors. (C) Quantitative PCR analysis of miR-488-3p expression in xenograft tumors. (D) Representative immunohistochemical staining of vimentin, N-cadherin, E-cadherin and Ki-67 expression in xenograft tumors. (E) Schematic representation of the MEIS1/miR-488-3p/ACVR1C axis in regulating laryngeal squamous cell carcinoma progression. The red and green arrows indicate upregulation and downregulation in LSCC, respectively. Scale bar, 50 mm. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; NC, negative control; MEIS1, myeloid ecotropic viral integration site 1; ACVR1C, activin A receptor type 1C.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: In Vivo, Injection, Real-time Polymerase Chain Reaction, Expressing, Immunohistochemical staining, Staining, Negative Control

Journal: bioRxiv

Article Title: Apical expansion of calvarial osteoblasts and suture patency is dependent on graded fibronectin cues

doi: 10.1101/2023.01.16.524278

Figure Lengend Snippet: (A) Immunofluorescence of unfolded collagen chains in E12.5 coronal sections. Immunofluorescence of collagen III (B), Laminin (C), RGB Trichrome staining for collagen (D) in E12.5 coronal sections. (E, F, G) Immunofluorescence of FN1 in coronal sections showing graded FN1 protein expression in cranial mesenchyme at E11.5-13.5. N≥3 controls; 3 mutants. (H) Schematic depicting the graded FN1 substrate towards the apex of the cranial mesenchyme. Nuclei were labeled with DAPI in blue. CHP, collagen hybridizing peptide; FN1, Fibronectin 1; fbp, frontal bone primordia. mn, meninges. Scale Bar= 100 μM.

Article Snippet: Following primary antibodies were used: Collagen hybridizing peptide (CHP) (Hwang et al., 2017; Jussila et al., 2021) (1: 250; 3Helix BIO60), rabbit anti-Fibronectin antibody (1: 250; Abcam ab2413; RRID:AB_2262874), rabbit anti-Fibulin 1 (1: 250; Abcam ab230994), rabbit anti-Collagen I (1: 250; Abcam ab270993; RRID:AB_2927551), Collagen III (Rockland Labs 1:100), rabbit anti-Laminin (1: 250; Millipore L9393), rabbit anti-Osx (1:2000 or 1:4000; Abcam ab209484; RRID:AB_2892207), rabbit anti-Caspase 3 (1: 250; Abcam ab13847; RRID:AB_443014), Goat anti-Runx 2 (1:100; R&D Systems AF2006-SP; RRID:AB_2184528) and mouse anti-GM130 (1:100; Thermo Fisher Scientific BDB610822; RRID:AB_398141).

Techniques: Immunofluorescence, Staining, Expressing, Labeling