Journal: Oncogene

Article Title: Inhibition of autophagy and tumor growth in colon cancer by miR-502

doi: 10.1038/onc.2012.167

Figure Lengend Snippet: (A) A putative miR-502 binding site exists in the 3’-UTR of Rab1B mRNA and two point mutations were generated in the binding site. Ectopic expression of miR-502 or siRNAs against Rab1B (siRab1B) in HCT116 and SW480 cells decreased Rab1B protein levels by (B) Western blot analysis and (C) mRNA level by real-time qRT-PCR. (D) Transfection of miR-502 inhibited firefly luciferase activity of pMIR-REPORT-3UTRRab1B (wt) and such inhibition was absent with mutations in the miR-502 binding site (mut). The negative miRNA was used as the negative control in all experiments. The impact of miR-502 and siRab1B on Rab1B expression was normalized and compared to those of negative miRNA (n=3, p < 0.001).

Article Snippet: 10 pmole of miR-502 or negative miRNA was transfected into cells together with 100ng of pMIR-REPORT-3UTRRab1B and 1ng of Renilla luciferase plasmid pRL-SV40 (Promega) by DharmaFect Duo (Dharmacon).

Techniques: Binding Assay, Generated, Expressing, Western Blot, Quantitative RT-PCR, Transfection, Luciferase, Activity Assay, Inhibition, Negative Control

Journal: Cells

Article Title: Hydrogen Peroxide Inhibits Hepatitis C Virus Replication by Downregulating Hepatitis C Virus Core Levels through E6-Associated Protein-Mediated Proteasomal Degradation

doi: 10.3390/cells13010062

Figure Lengend Snippet: H 2 O 2 downregulates HCV Core levels by upregulating E6AP levels in a p53-dependent manner. ( a – c ) Cells were transfected with the specified plasmids for 24 h and treated with H 2 O 2 for an extra 24 h, followed by Western blotting.

Article Snippet: The plasmid pCMVT N-HA-hE6AP, encoding the complete human HA-tagged E6AP , was purchased from Addgene (Watertown, MA, USA).

Techniques: Transfection, Western Blot

Journal: Cells

Article Title: Hydrogen Peroxide Inhibits Hepatitis C Virus Replication by Downregulating Hepatitis C Virus Core Levels through E6-Associated Protein-Mediated Proteasomal Degradation

doi: 10.3390/cells13010062

Figure Lengend Snippet: H 2 O 2 stimulates E6AP expression through promoter hypomethylation in the presence of p53 and HCV Core. Cells were transfected with the specified plasmids for 24 h and subsequently treated with H 2 O 2 for an extra 24 h. ( a , b ) DNMT activity from cells was determined ( n = 3). Levels of the indicated proteins were measured by Western blotting. ( c , d ) Methylation-specific PCR (MSP) was performed to determine whether the CpG sites in the E6AP promoter are unmethylated (U) or methylated (M). ( e ) Cells were treated with the specified concentration of 5-Aza-2′dC for 24 h before harvesting.

Article Snippet: The plasmid pCMVT N-HA-hE6AP, encoding the complete human HA-tagged E6AP , was purchased from Addgene (Watertown, MA, USA).

Techniques: Expressing, Transfection, Activity Assay, Western Blot, Methylation, Concentration Assay

Journal: Cells

Article Title: Hydrogen Peroxide Inhibits Hepatitis C Virus Replication by Downregulating Hepatitis C Virus Core Levels through E6-Associated Protein-Mediated Proteasomal Degradation

doi: 10.3390/cells13010062

Figure Lengend Snippet: H 2 O 2 triggers the E6AP-mediated ubiquitination and proteasomal degradation of HCV Core in a p53-dependent manner. ( a ) Cells prepared as in a,b were subjected to treatment with 50 μM cycloheximide (CHX) for the specified duration before harvesting. The quantification of each band was performed using Image J image-analysis software (NIH, USA) to determine the half-life (t 1/2 ) of HCV Core. The presented values represent the levels of HCV Core relative to the loading control (γ-tubulin). ( b ) Cells were transfected with the specified plasmids for 24 h and treated with H 2 O 2 for an extra 24 h. The transfection mixtures included the HA-Ub expression plasmid. Total HCV Core proteins in cell lysates were immunoprecipitated using an anti-HCV Core antibody and subsequently analyzed by Western blotting. The membranes were probed with antibodies against p53, HCV Core, E6AP, and HA to detect p53, HCV Core, E6AP, and HA-Ub-complexed HCV Core, respectively. Additionally, the input shows the levels of the specified proteins in the cell lysates. ( c ) Cells were either mock-treated or treated with MG132 for 4 h before harvesting.

Article Snippet: The plasmid pCMVT N-HA-hE6AP, encoding the complete human HA-tagged E6AP , was purchased from Addgene (Watertown, MA, USA).

Techniques: Software, Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot

Journal: Cells

Article Title: Hydrogen Peroxide Inhibits Hepatitis C Virus Replication by Downregulating Hepatitis C Virus Core Levels through E6-Associated Protein-Mediated Proteasomal Degradation

doi: 10.3390/cells13010062

Figure Lengend Snippet: H 2 O 2 inhibits HCV replication in vitro through the E6AP-mediated downregulation of HCV Core levels. ( a ) Cells were transfected with the designated plasmids for 24 h and infected with HCV for an extra 24 h in the absence or presence of H 2 O 2 , followed by Western blot analysis. ( b ) The levels of HCV particles in the supernatants, as prepared in ( a ), were determined using both q-RT-PCR ( n = 4) and conventional RT-PCR. ( c ) Cells cultured in a differentiation medium (Biopredic International) for 2 weeks were infected with HCV for 24 h and treated with H 2 O 2 for an extra 24 h, followed by Western blotting. ( d ) The levels of HCV particles in the supernatants prepared in ( c ) were determined using q-RT-PCR ( n = 3) and conventional RT-PCR. ( e ) Cells were transfected with the specified plasmids for 24 h and subsequently infected with HCV for an extra 24 h, either in the absence or presence of H 2 O 2 , followed by co-IP, as described in b.

Article Snippet: The plasmid pCMVT N-HA-hE6AP, encoding the complete human HA-tagged E6AP , was purchased from Addgene (Watertown, MA, USA).

Techniques: In Vitro, Transfection, Infection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Co-Immunoprecipitation Assay

Journal: Molecular Medicine Reports

Article Title: TNF-α treatment increases DKK1 protein levels in primary osteoblasts via upregulation of DKK1 mRNA levels and downregulation of miR-335-5p

doi: 10.3892/mmr.2020.11152

Figure Lengend Snippet: DKK1 mRNA levels in MC3T3-E1 cells and primary calvarial osteoblasts treated with TNF-α. (A) DKK1 mRNA levels in MC3T3-E1 cells and primary calvarial osteoblasts treated with 15 ng/ml TNF-α for 0, 12, 24 and 48 h. *P<0.05 vs. the 0-h group, n=3. (B) DKK1 mRNA levels in MC3T3-E1 cells and primary calvarial osteoblasts treated with 0, 15 and 50 ng/ml TNF-α for 48 h. *P<0.05 vs. the 0-ng/ml group, n=3. DKK1, Dickkopf WNT signaling pathway inhibitor 1; TNF-α, tumor necrosis factor-α.

Article Snippet: For transient transfection, MC3T3-E1 cells were cultured in 12-well plates at a density of 5×10 4 cells per ml overnight, and co-transfected with 900 ng pMIR-REPORT-DKK1 UTR and 100 ng pRL-TK Vector (Promega Corporation) using Lipofectamine ® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques:

Journal: Molecular Medicine Reports

Article Title: TNF-α treatment increases DKK1 protein levels in primary osteoblasts via upregulation of DKK1 mRNA levels and downregulation of miR-335-5p

doi: 10.3892/mmr.2020.11152

Figure Lengend Snippet: DKK1 protein levels in MC3T3-E1 cells and primary calvarial osteoblasts treated with TNF-α. (A) Protein levels of DKK1 in MC3T3-E1 cells and primary calvarial osteoblasts treated with 15 ng/ml TNF-α for 0, 12, 24 and 48 h. *P<0.05 vs. the 0-h group, n=3. (B) Protein levels of DKK1 in MC3T3-E1 cells and primary calvarial osteoblasts treated with 0, 15 and 50 ng/ml TNF-α for 48 h. *P<0.05 vs. the 0-ng/ml group, n=3. DKK1, Dickkopf WNT signaling pathway inhibitor 1; TNF-α, tumor necrosis factor-α.

Article Snippet: For transient transfection, MC3T3-E1 cells were cultured in 12-well plates at a density of 5×10 4 cells per ml overnight, and co-transfected with 900 ng pMIR-REPORT-DKK1 UTR and 100 ng pRL-TK Vector (Promega Corporation) using Lipofectamine ® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques:

Journal: Molecular Medicine Reports

Article Title: TNF-α treatment increases DKK1 protein levels in primary osteoblasts via upregulation of DKK1 mRNA levels and downregulation of miR-335-5p

doi: 10.3892/mmr.2020.11152

Figure Lengend Snippet: Role of the NF-κB signaling pathway in mediating TNF-α-regulated expression of DKK1 in MC3T3-E1 cells and primary calvarial osteoblasts. MC3T3-E1 cells and primary calvarial osteoblasts were treated with 15 ng/ml TNF-α and/or 1 µM BAY 11-7082 for 48 h, and the untreated cells served as controls. The DKK1 (A) mRNA levels and (B) protein levels in these cells were determined. *P<0.05 vs. the Control group; # P<0.05 vs. the TNF-α group; & P<0.05 vs. the TNF-α + BAY 11-7082 group, n=3. TNF-α, tumor necrosis factor-α; DKK1, Dickkopf WNT signaling pathway inhibitor 1.

Article Snippet: For transient transfection, MC3T3-E1 cells were cultured in 12-well plates at a density of 5×10 4 cells per ml overnight, and co-transfected with 900 ng pMIR-REPORT-DKK1 UTR and 100 ng pRL-TK Vector (Promega Corporation) using Lipofectamine ® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: TNF-α treatment increases DKK1 protein levels in primary osteoblasts via upregulation of DKK1 mRNA levels and downregulation of miR-335-5p

doi: 10.3892/mmr.2020.11152

Figure Lengend Snippet: TNF-α treatment exhibits no effects on the inhibitory action of miR-335-5p via targeting of DKK1 3′UTR. MC3T3-E1 cells were co-transfected with pMIR-REPORT-DKK1 UTR and pRL-TK Vector, and cells co-transfected with pMIR-REPORT and pRL-TK served as controls. (A) Transfected MC3T3-E1 cells were treated with 15 ng/ml TNF-α for 0, 12, 24 and 48 h, and the luciferase levels were determined. *P<0.05 vs. the pMIR-REPORT group, n=3. (B) Transfected MC3T3-E1 cells were treated with 0, 15 and 50 ng/ml TNF-α for 48 h, and the luciferase levels were determined. *P<0.05, vs. the pMIR-REPORT group, n=3. (C) mmu-miR-335-5p mimic or mirVana™ miRNA Mimic Negative Control #1 were transfected into the MC3T3-E1 cells concurrently, and the luciferase levels were determined. The transfected cells were treated with or without 15 ng/ml TNF-α for 48 h. The luciferase levels were determined. *P<0.05, vs. the pMIR-REPORT group, n=3. mimic NC= miRNA Mimic negative Control. (D) Transfected MC3T3-E1 cells were treated with or without 15 ng/ml TNF-α and/or 1 µM BAY 11-7082 for 48 h. *P<0.05 vs. the pMIR-REPORT group, n=3. TNF-α, tumor necrosis factor-α; miR, microRNA; DKK1, Dickkopf WNT signaling pathway inhibitor 1.

Article Snippet: For transient transfection, MC3T3-E1 cells were cultured in 12-well plates at a density of 5×10 4 cells per ml overnight, and co-transfected with 900 ng pMIR-REPORT-DKK1 UTR and 100 ng pRL-TK Vector (Promega Corporation) using Lipofectamine ® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.).

Techniques: Transfection, Plasmid Preparation, Luciferase, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: LINC00312 may act as an miR-4765 sponge to regulate FOXK2 and SFRP1 as predicted using the database. (A) Transfection efficiency of LINC00312 OE plasmids and sh-LINC00312. (B) miR-4765 expression after transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. (C) FOXK2 and SFRP1 protein expression after transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. (D) FOXK2 and SFRP1 mRNA expression in the bone tissues of diabetic mice with OP after adenovirus transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. (E) FOXK2 and SFRP1 protein expression determined using immunohistochemical staining in the bone tissues of diabetic mice with OP after adenovirus transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. Data are presented as the mean ± standard deviation (n=3). * P<0.05 and ** P<0.01. miR, microRNA; NC, negative control; OE, overexpression; sh-, short hairpin; OP, osteoporosis.

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Transfection, Expressing, Immunohistochemical staining, Staining, Standard Deviation, Negative Control, Over Expression

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: LINC00312 can be directly combined with miR-4765. (A) Binding sites for LINC00312 and miR-4765. (B) Luciferase activity in 293T cells co-transfected with miR-4765 mimics and LINC00312 WT or MUT 3' region. (C) Viability of hFOB 1.19 cells after co-transfection with LINC00312 OE plasmids and miR-4765 mimics and the corresponding NCs. Cell viability was expressed as OD values. (D) Cleaved caspase-3 protein expression after co-transfection with LINC00312 OE plasmids and miR-4765 mimics and the corresponding NCs. (E) Viability of hFOB 1.19 cells after co-transfection with LINC00312 OE plasmids and sh-FOXK2 and the corresponding NCs. Cell viability was expressed as OD values. (F) Cleaved caspase-3 protein expression after co-transfection with LINC00312 OE plasmids and sh-FOXK2 and the corresponding NCs. (G) Viability of hFOB 1.19 cells after co-transfection with LINC00312 OE plasmids and sh-SFRP1 and the corresponding NCs. Cell viability was expressed as OD values. (H) Cleaved caspase-3 protein expression after co-transfection with LINC00312 OE plasmids and sh-SFRP1 and the corresponding NCs. Data are presented as the mean ± standard deviation (n=3). * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; WT, wild type; MUT, mutant; OD, optical density; NC, negative control; OE, overexpression; sh-, short hairpin.

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Cotransfection, Expressing, Standard Deviation, Mutagenesis, Negative Control, Over Expression

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: LINC00312 can promote hFOB 1.19 cells apoptosis. (A) Viability of hFOB 1.19 cells after transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. Cell viability was expressed as OD values. (B) Cleaved caspase-3 protein expression after transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. (C) Apoptosis levels determined via Annexin V staining after transfection with LINC00312 OE plasmids, sh-LINC00312, and the corresponding NCs. (D) LINC00312 expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). Data are presented as the mean ± standard deviation (n=3). * P<0.05 and ** P<0.01. OD, optical density; NC, negative control; OE, overexpression; sh-, short hairpin.

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Transfection, Expressing, Staining, Standard Deviation, Negative Control, Over Expression

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: METTL3 reduces the expression level of LINC00312 by increasing its methylation level. (A) SRAMP was used to predict the possible m6A modification locations of LINC00312. (B) Relative m6A level of hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l) determined via m6A colorimetric analysis. (C) METTL3 mRNA expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). (D) METTL3 protein expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). (E) Transfection efficiency of METTL3 OE plasmids and sh-METTL3. METTL3 mRNA expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (F) Transfection efficiency of METTL3 OE plasmids and sh-METTL3; METTL3 protein expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (G) LINC00312 expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (H) miR-4765 expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (I) FOXK2 and SFRP1 protein expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (J) Viability of hFOB 1.19 cells after transfection with METTL3 OE plasmid, sh-METTL3, and the corresponding NCs. Cell viability was expressed as OD values. (K) Cleaved caspase-3 protein expression after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (L) Apoptosis levels determined via Annexin V staining after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (M) m6A modification locations of LINC00312. (N) Luciferase activity in 293T cells co-transfected with METTL3 OE plasmids and LINC00312 WT or MUT 3'-region. (O) Relative enrichment of METTL3 in LINC00312 after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (P) Relative enrichment of m6A in LINC00312 under various glucose concentrations (1 or 4.5 g/l) determined via methylated RNA immunoprecipitation-quantitative PCR assays. (Q) Relative enrichment of m6A in LINC00312 after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (R) METTL3 protein expression determined via IHC staining in the bone tissues of diabetic mice with OP and non-diabetic mice. (S) FOXK2 and SFRP1 mRNA expression in the bone tissues of diabetic mice with OP after adenovirus transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (T) FOXK2 and SFRP1 protein expression determined via IHC staining in the bone tissues of diabetic mice with OP after adenovirus transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. Data are presented as the mean ± standard deviation (n=3). * P<0.05 and ** P<0.01. OE, overexpression; sh-, short hairpin; WT, wild type; MUT, mutant; NC, negative control; OD, optical density; miR, microRNA; OP, osteoporosis; IHC, immunohistochemical.

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Expressing, Methylation, Modification, Transfection, Plasmid Preparation, Staining, Luciferase, Activity Assay, RNA Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunohistochemistry, Standard Deviation, Over Expression, Mutagenesis, Negative Control, Immunohistochemical staining

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: YTHDF2 participates in increasing the methylation level of LINC00312. (A) YTHDF2 mRNA expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). (B) YTHDF2 protein expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). (C) Transfection efficiency of YTHDF2 OE plasmids and sh-YTHDF2. YTHDF2 mRNA expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (D) Transfection efficiency of YTHDF2 OE plasmids and sh-YTHDF2. YTHDF2 protein expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (E) LINC00312 expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2 and the corresponding NCs. (F) miR-4765 expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (G) FOXK2 and SFRP1 protein expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (H) Viability of hFOB 1.19 cells after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. Cell viability was expressed as OD values. (I) Cleaved caspase-3 protein expression after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (J) Apoptosis level determined via Annexin V staining after transfection with YTHDF2 OE plasmids, sh-YTHDF2, and the corresponding NCs. (K) m6A modification locations of LINC00312. (L) Luciferase activity in 293T cells co-transfected with YTHDF2 OE plasmids and LINC00312 WT or MUT 3'-region. (M) Relative enrichment of YTHDF2 in LINC00312 after transfection with METTL3 OE plasmids, sh-METTL3, and the corresponding NCs. (N) YTHDF2 protein expression determined via immunohistochemical staining in the bone tissues of diabetic mice with osteoporosis and non-diabetic mice. Data are presented as the mean ± standard deviation (n=3). * P<0.05 and ** P<0.01. WT, wild type; MUT, mutant; NC, negative control; OD, optical density; OE, overexpression; sh-, short hairpin; miR, microRNA.

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Methylation, Expressing, Transfection, Staining, Modification, Luciferase, Activity Assay, Immunohistochemical staining, Standard Deviation, Mutagenesis, Negative Control, Over Expression

Journal: International Journal of Molecular Medicine

Article Title: METTL3 inhibits LINC00312 to suppress osteoporosis progression in a YTHDF2-dependent manner

doi: 10.3892/ijmm.2025.5699

Figure Lengend Snippet: Graphic abstract. LINC00312 increases apoptosis of hFOB 1.19 cells by targeting the miR-4765-FOXK2/SFRP1 axis, which is m6A modified by METTL3 in a YTHDF2-dependent manner. miR, microRNA. The image was created using www.Figdraw.com .

Article Snippet: 293T cells (Chinese Academy of Sciences) were seeded into 24-well plates and cultured overnight, followed by transfection with wild- and mutant-type LINC00312 plasmids (0.1 μ g pMIR-REPORT-wild-type-LINC00312 or pMIR-RE PORT-mutant-type-LINC00312 plasmids per well) (Shanghai GeneChem Co., Ltd.) using Roche X-tremeGENE HP (cat. no. 06366236001; Roche Diagnostics) according to the manufacturer's instructions.

Techniques: Modification