Journal: BMC Developmental Biology

Article Title: BMP-SMAD signalling output is highly regionalized in cardiovascular and lymphatic endothelial networks

doi: 10.1186/s12861-016-0133-x

Figure Lengend Snippet: The GFP reporter protein faithfully recapitulates the transcriptional activation of the BRE :: gfp transgene. In situ hybridisation for gfp mRNA ( red ) and direct GFP fluorescence ( green ) on cryosections through the cardinal vein ( a – d ) and the ventricular trabeculae ( e – g ) at E11.5. DAPI is used to stain nuclei. The boxed area in ( a ) is enlarged in panels ( b – d ). The corresponding low magnification view of the trabeculae in the ventricle is not provided because of insufficient intensity of the ISH signal. White arrowheads indicate ECs double positive for gfp mRNA and direct GFP fluorescence. The red and green arrows depict ECs only positive for gfp mRNA or GFP fluorescence respectively. Scale bars: 100 μm ( a ); 50 μm ( b – d ); 10 μm ( e – g )

Article Snippet: The GFP fluorescent in situ hybridisation (ISH) probe was custom designed with the probe designer tool from Stellaris (LGC biosearch technologies).

Techniques: Activation Assay, In Situ, Hybridization, Fluorescence, Staining

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: Relationship between LINC02038 expression and clinical characteristics in patients with colorectal cancer (n=68).

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: LINC02038 is downregulated in CRC. (A) Heatmap of lncRNAs. lncRNAs with an FC ≥5 and P<0.05 were considered down- or upregulated. Downregulation is represented by a blue scale, while upregulation is represented by a red scale. (B) lncRNA volcano diagram of CRC vs. non-tumorous tissues. Upregulated genes are indicated in red and downregulated genes are indicated in green (defined by FC ≥1 and P<0.05). Genes in black were differentially expressed by FC <1. (C) Expression of LINC02038 based on pan-cancer analysis utilizing the TCGA and GTEx databases. LINC02038 expression was lower in COAD tissues than in normal colorectal tissues. (D and E) The expression of LINC02038 in CRC and normal colorectal tissues based on data obtained from GTEx and TCGA datasets. (F) The relationship between the LINC02038 and OS in the obtained datasets. (G) Bubble plots showing the top 21 dysregulated KEGG pathways in terms of genes associated with LINC02038 in CRC. * P<0.05, *** P<0.001. CRC, colorectal cancer; lncRNA, long non-coding RNA; TCGA, The Cancer Genome Atlas; GTEx, Genotype-Tissue Expression project; KEGG, Kyoto Encyclopedia of Genes and Genomes; COAD, colon adenocarcinoma; OS, overall survival; ns, no significance.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: LINC02038 suppresses CRC cell growth, motility and invasion while inducing apoptosis. (A) LINC02038 expression in 68 paired CRC and normal colorectal tissue. (B and C) LINC02038 expression in several different CRC cell lines. (D) LINC02038 was successfully overexpressed in CRC cells transfected with pcDNA3.1-LINC02038. (E) CCK-8 assay results showed the proliferative effects of LINC02038 overexpression on CRC cells. (F) Overexpression of LINC02038 affected CRC cell cycle regulation. (G) CRC proliferation was detected using colony formation assays following LINC02038 overexpression. (H) Overexpression of LINC02038 promoted apoptosis in a fraction of the cells as measured by flow cytometry. Transwell assays showing CRC (I) migration and (J) invasion following LINC02038 overexpression. Magnification, ×200. * P<0.05, ** P<0.01 vs. normal tissues, NCM460, or empty vector; CRC, colorectal cancer.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Expressing, Transfection, CCK-8 Assay, Over Expression, Flow Cytometry, Migration, Plasmid Preparation

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: LINC02038 sponges miR-552-5p in the CRC cells. (A) LINC08038 localized to the cytoplasm (red) and nucleus (blue) in LoVo and SW620 cells using FISH test. (B) The predicted binding site between LINC02038 and miR-552-5p. (C) Using data obtained from TCGA-COAD dataset, it was shown there was a negative association between LINC02038 and miR-552-5p expression in CRC tissues. (D) TCGA-COAD database analysis confirmed that miR-552-5p expression was higher in CRC tissues than in matched and unpaired normal tissues. (E) RT-qPCR analysis of miR-552-5p expression in CRC and normal tissues. (F) RT-qPCR detection of miR-552-5p expression in CRC cells. (G) miR-552-5p expression in LoVo and SW620 cells transfected with LINC02038 overexpression or an empty vector was determined using RT-qPCR. (H) The relative luciferase activity in LoVo and SW620 cells co-transfected with LINC02038 WT or MUT and miR-552-5p mimics. (I) RIP assays were used to examine LINC02038 enrichment in LoVo and SW620 cells co-transfected with LINC0203 and miR-552-5p mimic. (J) The relationship between miR-552-5p and overall survival in CRC patients in TCGA-COAD dataset. ** P<0.01, *** P<0.001. miR, microRNA; CRC, colorectal cancer; FISH, Fluorescence in situ hybridization; TCGA, The Cancer Genome Atlas; COAD, colon adenocarcinoma; RT-qPCR, reverse transcription quantitative PCR; MUT, mutant; WT, wild-type; Ago2, argonaute 2; RIP, RNA immunoprecipitation.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Luciferase, Activity Assay, Fluorescence, In Situ Hybridization, Real-time Polymerase Chain Reaction, Mutagenesis, Immunoprecipitation

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: LINC02038 exerts its function by inhibiting miR-552-5p in CRC cells. (A) miR-552-5p expression was determined using RT-qPCR after transfection with miR-552-5p mimic, LINC02038 overexpression + miR-552-5p mimic or the control. (B) CCK-8 and (C) colony formation assays were used to assess the proliferation of transfected cells. (D and E) Flow cytometry was used to identify the cell cycle distribution and apoptosis of SW620 and LoVo cells transfected with miR-552-5p mimic, LINC02038 overexpression + miR-552-5p mimic or the control. LINC02038 overexpression resulted in G 1 phase cell cycle arrest and increased rates of apoptosis and partially reversed the effects of miR-552-5p mimic transfection in CRC cells. (F) Migration and (G) invasion rates were determined using Transwell assays in CRC cells co-transfected with LINC02038 overexpression vector and miR-552-5p mimics as well as the corresponding controls. * P<0.05, ** P<0.01 vs. NC mimic; miR, microRNA; CRC, colorectal cancer; RT-qPCR, reverse transcription quantitative PCR; NC, negative control.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Over Expression, CCK-8 Assay, Flow Cytometry, Migration, Plasmid Preparation, Real-time Polymerase Chain Reaction, Negative Control

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: LINC02038 inhibits CRC progression by regulating FAM172A. (A) The knockdown efficiency of siRNAs targeting FAM172A was validated using RT-qPCR. (B) FAM172A expression was examined using western blotting in the FAM172A knockdown cells and/or LINC02038 overexpressing cells. (C) FAM172A mRNA levels are shown in SW620 and LoVo cells co-transfected with siFAM172A and LINC02038 overexpression vector. (D and E) CCK-8 and colony formation assays were used to evaluate cell viability. (F) Flow cytometry was used to examine the cell cycle distribution. (G) Apoptosis was detected in SW620 and LoVo cells using Annexin V/PI double-staining by fluorescence activated cell sorting. Transwell assays were used to evaluate (H) migration and (I) invasion of SW620 and LoVo cells. Scale bars, 100 µ m. (J) The protein expression levels of AKT, p-AKT and PI3K were analyzed by western blotting in SW620 and LoVo cells. Data are presented as the mean ± SD of three repeats. * P<0.05, ** P<0.01. CRC, colorectal cancer; siRNA, small interfering RNA; RT-qPCR, reverse transcription quantitative PCR; p-, phosphorylated.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Transfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Flow Cytometry, Double Staining, Fluorescence, FACS, Migration, Small Interfering RNA, Real-time Polymerase Chain Reaction

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: In vivo , LINC02038 inhibits CRC tumorigenesis. (A) Images of the CRC tissues following subcutaneous injection of LoVo cells with LINC02038 overexpression or pcDNA3.1/Control empty vector. The (B) volume and (C) weight of xenograft tumors were measured. (D) Representative images of IHC-stained CRC tissues. Magnification, ×100 and ×200. Data are presented as the mean ± SD of three repeats. ** P<0.01. CRC, colorectal cancer; IHC, immunohistochemistry.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: In Vivo, Injection, Over Expression, Plasmid Preparation, Staining, Immunohistochemistry

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: METTL3 knockdown reduces the stability of LINC02038 via an m 6 A-YTHDF2-dependent mechanism. (A) SRAMP was used to predict the possible m 6 A modification locations of LINC02038. (B) The enrichment of m 6 A-modified LINC02038 was measured using a MeRIP-qPCR assay. (C) Using RT-qPCR and western blotting, METTL3 expression was determined in CRC cells following METTL3 knockdown. (D) Correlation analysis of METTL3 and LINC02038 mRNA on data obtained from TCGA. (E) LINC02038 expression in METTL3-deficient LoVo and SW620 cells was detected using RT-qPCR. (F) shMETTL3-mediated LINC02038 m 6 A modifications were demonstrated in LoVo and SW620 cells. (G) The interaction between METTL3, METTL14, FTO and YTHDF2. (H) YTHDF2 expression of LoVo and SW620 cells transfected with shYTHDF2 were detected using RT-qPCR and western blotting. (I) LINC02038 expression was determined using RT-qPCR in YTHDF2-deficient LoVo and SW620 cells. (J) The expression of LINC02038 was detected following treatment with Actinomycin D (5 mg/ml) in SW620 cells using RT-qPCR at the indicated time points. Data are presented as the mean ± SD of three repeats. * P<0.05, ** P<0.01. m 6 A, N6-methyladenosine; MeRIP, methylated RNA immunoprecipitation; qPCR, quantitative PCR; RT-qPCR, reverse transcription qPCR; TCGA, The Cancer Genome Atlas; shRNA, short hairpin RNA.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques: Modification, Quantitative RT-PCR, Western Blot, Expressing, Transfection, Methylation, Immunoprecipitation, Real-time Polymerase Chain Reaction, shRNA

Journal: International Journal of Oncology

Article Title: m 6 A-mediated LINC02038 inhibits colorectal cancer progression via regulation of the FAM172A/PI3K/AKT pathway via competitive binding with miR-552-5p

doi: 10.3892/ijo.2023.5529

Figure Lengend Snippet: Graphic illustrating the m 6 A-dependent effects of the LINC02038/miR-552-5p/FAM172A/PI3K/AKT regulatory loop on CRC growth. m 6 A, N6-methyladenosine; CRC, colorectal cancer; miR, microRNA.

Article Snippet: GFP-labeled LINC02038 probes were supplied by Shanghai GenePharma Co., Ltd. A FISH kit (Shanghai GenePharma Co., Ltd.) was used to perform the hybridization.

Techniques:

Journal: Scientific Reports

Article Title: Paeoniflorin inhibits angiogenesis in multiple myeloma by decreasing the MEF2A level to downregulate the expression of lncRNA MALAT1 within exosomes

doi: 10.1038/s41598-025-30101-6

Figure Lengend Snippet: The clinical significance of MALAT1 and its correlation with angiogenesis. (A) LncRNAs were distributed on 22 autosomes and sex chromosomes (X, Y). (B) The lncRNA categories to which each of the 2052 circRNAs belongs. (C) Distribution of the lncRNAs based on the predicted sequence length. (D) Volcano plot of DEGs from GEO5900 dataset. (E) The top 10 expression lncRNAs in U266 exosomes. (F) KM curve analysis comparing MALAT1 high-risk and low-risk groups. ( P < 0.0001). (G) The accuracy of MALAT1 in predicting the death outcome of MM patients. (H , I) Analysis of MALAT1 FISH in high-risk and low-risk groups of MM patients. (J) CD31 and VEGFA IHC images of MM patients’ BM. (K) Analysis of MVD and VEGFA level in high-risk and low-risk groups. (L) MALAT1 expression positively correlated with MVD and VEGFA levels. Three independent experiments were performed. * P < 0.05, ** P < 0.01.

Article Snippet: GFP-labeled MALAT1 probes, synthesized by Beijing Dingguo Changsheng Biotech Co., Ltd., were utilized for the FISH analysis.

Techniques: Sequencing, Expressing

Journal: Scientific Reports

Article Title: Paeoniflorin inhibits angiogenesis in multiple myeloma by decreasing the MEF2A level to downregulate the expression of lncRNA MALAT1 within exosomes

doi: 10.1038/s41598-025-30101-6

Figure Lengend Snippet: Mechanistic study of MALAT1 derived from U266 enhance angiogenesis in HUVECs. (A) The shapes and sizes of serum exosomes were observed by TEM. (B) The average diameter and concentration of exosomes were calculated by NTA. (C) The intercellular trafficking of MALAT1 is facilitated by exosomes, which are taken up by HUVEC cells. This process was observed using isolated exosomes labeled with PKH26 dye and MALAT1 labeled with GFP. (D) The cell viability assays were performed on HUVEC cells, both with and without MALAT1 knockdown, following co-culture with exosomes. (E) MALAT1 level was tested by PCR. (F) VEGFA level was tested by PCR. (G) VEGFA level was tested by ELISA. (H) Tube formation assays were performed on HUVEC cells, both with and without MALAT1 knockdown, following co-culture with exosomes. (I) The ceRNA network diagram illustrating the interaction between MALAT1, miRNAs, and target mRNA genes. (J) The direct interaction between miR-17 and MALAT1 as well as VEGFA was confirmed through dual-luciferase reporter assays.Three independent experiments were performed. * P < 0.05, ** P < 0.01.

Article Snippet: GFP-labeled MALAT1 probes, synthesized by Beijing Dingguo Changsheng Biotech Co., Ltd., were utilized for the FISH analysis.

Techniques: Derivative Assay, Concentration Assay, Isolation, Labeling, Knockdown, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Luciferase

Journal: Scientific Reports

Article Title: Paeoniflorin inhibits angiogenesis in multiple myeloma by decreasing the MEF2A level to downregulate the expression of lncRNA MALAT1 within exosomes

doi: 10.1038/s41598-025-30101-6

Figure Lengend Snippet: PF inhibit angiogenesis by decrease the MALAT1 level of exosomes. (A) The MALAT1 level was assessed using PCR in different dosage groups treated with PF, the medium-dose PF treated group was selected for further investigation based on the level of MALAT1. (B) The cell viability assays were conducted on HUVEC cells, following co-culture with exosomes which derived from U266 cells after treated with or without a medium dose of PF. (C , D) Tube formation assays were performed on HUVEC cells. (E , F) VEGFA level was tested by PCR or ELISA. Three independent experiments were performed. * P < 0.05, ** P < 0.01.

Article Snippet: GFP-labeled MALAT1 probes, synthesized by Beijing Dingguo Changsheng Biotech Co., Ltd., were utilized for the FISH analysis.

Techniques: Co-Culture Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Paeoniflorin inhibits angiogenesis in multiple myeloma by decreasing the MEF2A level to downregulate the expression of lncRNA MALAT1 within exosomes

doi: 10.1038/s41598-025-30101-6

Figure Lengend Snippet: Effect of PF on mice model. (A) Compared to the EXO group, the tumor volume was significantly reduced when injected with exosomes pretreated with medium and high dose PF. (B) MALAT1 level was tested by PCR. (C) HE staining of the excised tumors from different experimental groups. (D) The tumor weight was analyzed in different experimental groups. (E , F) VEGFA level was tested by PCR or ELISA. (G , H) The evaluation of Ki67, CD31, and VEGFA IHC images in tumor tissues. (I) Analysis of MVD level in different experimental groups.Three independent experiments were performed. * P < 0.05, ** P < 0.01.

Article Snippet: GFP-labeled MALAT1 probes, synthesized by Beijing Dingguo Changsheng Biotech Co., Ltd., were utilized for the FISH analysis.

Techniques: Injection, Staining, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Paeoniflorin inhibits angiogenesis in multiple myeloma by decreasing the MEF2A level to downregulate the expression of lncRNA MALAT1 within exosomes

doi: 10.1038/s41598-025-30101-6

Figure Lengend Snippet: The mechanism PF on MALAT1. (A) The structure of PF. (B) The Venn plot of intersection genes of MALAT1 pull down gene, TF gene and PF target genes. (C) The binding site between MALAT1 and MEF2A was displayed. (D) The docking sites of PF between MEF2A. (E) WB analysis of MEF2A level in U266 cells treated with or without PF. (F) MALAT1 level in U266 cells treated with or without PF.Three independent experiments were performed. * P < 0.05, ** P < 0.01.

Article Snippet: GFP-labeled MALAT1 probes, synthesized by Beijing Dingguo Changsheng Biotech Co., Ltd., were utilized for the FISH analysis.

Techniques: Binding Assay