Journal: Cells

Article Title: Human Extravillous Trophoblasts Require SRC-2 for Sustained Viability, Migration, and Invasion

doi: 10.3390/cells14131024

Figure Lengend Snippet: Dysregulated expression of WNT signaling factors in HTR-8/SVneo cells following SRC-2 knockdown: ( A ) Heat map displays marked changes in the expression of genes associated with WNT signaling between the NT siRNA and SRC-2 siRNA replicate groups. While WNT signaling inhibitors, Dickkopf WNT signaling pathway inhibitor 1 ( DKK1 ) and secreted frizzled related protein 1 ( SFRP1 ), are upregulated in the SRC-2 siRNA-treated group, the expression levels of Wnt family member 2 ( WNT2 ), WNT4 , WNT6 , WNT8B , and WNT9A are significantly reduced. ( B ) Confirmation by qRT-PCR of the above-described gene expression changes between the NT siRNA and SRC-2 siRNA-treated groups is shown. An explanation of the average fold change (FC ± standard deviation) calculation is described in the . Results are representative of three independent experiments; * p -value < 0.05; ** p -value < 0.01, and *** p value < 0.001.

Article Snippet: SFRP1 , 6422 , Hs00610060_m1.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Gene Expression, Standard Deviation

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: FOXK2 and SFRP1 are potential target genes of miR-4765 predicted using databases. (A) Venn diagram of target genes overlapping with miR-4765 from three different databases (TargetScan, miRDB and DIANA). (B) FOXK2 mRNA expression after transfection with miR-4765 mimics, inhibitors, and the corresponding NCs. (C) SFRP1 mRNA expression after transfection with miR-4765 mimics, inhibitors, and the corresponding NCs. (D) FOXK2 and SFRP1 protein expression after transfection with miR-4765 mimics, inhibitors, and corresponding NCs. (E) Transfection efficiency of FOXK2 and SFRP1 OE plasmids, sh-FOXK2 and sh-SFRP1, and the corresponding NCs. (F) Viability of hFOB 1.19 cells after transfection with FOXK2 OE plasmid, sh-FOXK2, and the corresponding NCs. Cell viability was expressed as OD values. (G) Cleaved caspase-3 protein expression after transfection with FOXK2 OE plasmid, sh-FOXK2, and the corresponding NCs. (H) Viability of hFOB 1.19 cells after transfection with SFRP1 OE plasmid, sh-SFRP1, and the corresponding NCs. Cell viability was expressed as OD values. (I) Cleaved caspase-3 protein expression after transfection with SFRP1 OE plasmid, sh-SFRP1, and the corresponding NCs. (J) FOXK2 and SFRP1 expression in hFOB 1.19 cells under various glucose concentrations (1 or 4.5 g/l). (K) FOXK2 and SFRP1 mRNA expression in the bone tissues of control and diabetic mice. (L) FOXK2 and SFRP1 protein expression determined via immunohistochemical staining in the bone tissues of control and diabetic mice with OP. (M) Micro-computed tomography of the knee joint in control and diabetic mice with OP. Data are presented as the mean ± SD (n=3). * P<0.05 and ** P<0.01. miR, microRNA; NC, negative control; sh-, short hairpin; OD, optical density; OE, overexpression; OP, osteoporosis.

Article Snippet: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Immunohistochemical staining, Staining, Micro-CT, 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: miR-4765 can be directly combined with the 3'-UTR of FOXK2 and SFRP1. (A) Binding sites for miR-4765 and FOXK2, miR-4765 and SFRP1. (B) Luciferase activity in 293T cells co-transfected with miR-4765 mimic and FOXK2 WT or MUT 3'-UTR. (C) Luciferase activity in 293T cells co-transfected with miR-4765 mimic and SFRP1 WT or MUT 3'-UTR. (D) Viability of hFOB 1.19 cells after co-transfection with FOXK2 OE plasmids and miR-4765 mimics and the corresponding NCs. Cell viability was expressed as OD values. (E) Cleaved caspase-3 protein expression after co-transfection with FOXK2 OE plasmids and miR-4765 mimics and the corresponding NCs. (F) Viability of hFOB 1.19 cells after co-transfection with SFRP1 OE plasmids and miR-4765 mimics and the corresponding NCs. Cell viability was expressed as OD values. (G) Cleaved caspase-3 protein expression after co-transfection with SFRP1 OE plasmids and miR-4765 mimics 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; UTR, untranslated region; WT, wild type; MUT, mutant; OD, optical density; NC, negative control; OE, overexpression.

Article Snippet: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

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 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: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

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: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

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: 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: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

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: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

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: After washing with PBS, the sections were incubated for 30 min in 5% BSA at room temperature (20-25°C) and then incubated overnight in primary rabbit polyclonal anti-FOXK2 (cat. no. 30660-1-AP; Proteintech Group, Inc.) and anti-SFRP1 antibody (cat. no. 26460-1-AP; Proteintech Group, Inc.) at 4°C overnight.

Techniques: Modification