ythdf3 knockdown  (Obio Technology Corp Ltd)

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: YTHDF3 is overexpressed in breast cancer tissues and cells. (A) YTHDF3 expression in breast cancer tissues and adjacent normal tissues in TCGA database; (B) YTHDF3 expression in different types of breast cancer in TCGA database; (C) Kaplan–Meier curves of OS in the TCGA dataset of all breast cancer patients with high and low YTHDF3 expression; (D) YTHDF3 expression in paired breast cancer tissues and adjacent normal tissues; (E) YTHDF3 expression of breast cancer and para‐cancerous tissues was evaluated by immunohistochemistry, ×100; (F) YTHDF3 protein expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453; (G) Relative YTHDF3 mRNA expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: Expressing, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: Silence of YTHDF3 restrain cell proliferation and migration of breast cancer cells and arrested the cell cycle. (A) Relative expression of YTHDF3 mRNA and protein after transfection of shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (B) the growth ability of MDA‐MB‐231 and MCF‐7 cells transfected with shNC and shYTHDF3 was detected by CCK‐8 assay; (C) colony formation assay was used to detect the proliferation ability of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups; (D) the cell cycle distribution of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups was detected by flow cytometry; (E) expression of cyclin D1 and CDK4 in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (F, G) transwell invasion assay was used to detect the migration ability of breast cancer cells in the shNC group and the shYTHDF3 group in MCF‐7 and MDA‐MB‐231 cells; (H) the expression of vimentin, MMP9, E‐cadherin, N‐cadherin and snail in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: Migration, Expressing, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry, Transwell Invasion Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: Silencing YTHDF3 caused a decrease in lactate levels and affected glycolysis. (A) Lactate levels in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) extracellular acidification rate (ECAR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (C) oxygen consumption rate (OCR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (D) relative expression levels of hexokinase‐1 (HK1), hexokinase‐2 (HK2), PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (E) protein expression levels of PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; and (F) the correlation between YTHDF3 and PFKL, PKM, LDHA and LDHB was predicted using the GEPIA website.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: YTHDF3 regulates the glycolysis level of breast cancer through the mTOR–HIF1α–LDHA axis. (A) Relative mRNA expression of HIF1α in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) the correlation between YTHDF3 and HIF1α was predicted using the GEPIA website; (C) expression of key proteins in the mTOR–HIF1α–LDHA axis in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (D) after adding activator MHY1445, the growth ability of cancer cells in shNC and shYTHDF3 groups was detected by CCK‐8 assay; (E) the lactate level of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485; and (F) expression of proteins of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: Expressing, CCK-8 Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: YTHDF3 regulates breast cancer development through the mTOR–HIF1α–LDHA axis in vivo. (A) Tumour volumes in shNC and shYTHDF3 groups on the indicated days; (B) representative tumour images and (C) tumour weights in shNC and shYTHDF3 groups on day 63; (D) mRNA and (E) protein expression levels of YTHDF3, HIF1α and LDHA in the tumours in shNC and shYTHDF3 groups; (F) Immunohistochemical images of YTHDF3, Ki67, HIF1α and LDHA in tumours in the shNC and shYTHDF3 groups, ×100.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: In Vivo, Expressing, Immunohistochemical staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

doi: 10.1111/jcmm.71105

Figure Lengend Snippet: YTHDF3 is associated with prognosis in several cancers. (A) Column nomogram was constructed using YTHDF3 expression levels, age, T stage, N stage, M stage and tumour stage data of breast cancer patients; (B–E) survival analysis software package was used to analyse the survival prognosis difference between patients with high and low YTHDF3 expression in BRCA, DLBC, THCA and UVM.

Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

Techniques: Construct, Expressing, Software

Journal: Frontiers in Neurology

Article Title: Identification of m1A/m6A/m5C/m7G-related genes and clusters associated with neuropathic pain

doi: 10.3389/fneur.2026.1592545

Figure Lengend Snippet: Validation of expression of the six m1A/m6A/m5C/m7G-related DEGs ( FTO , METTL3 , NSUN2 , YTHDF3 , WDR4 , and EIF4E ) in rats after 3, 7, and 14 days of SNL surgery. n = 3 per group. SNL, spinal nerve ligation. (A) qPCR results. (B) Western blot analysis results. * p < 0.05.

Article Snippet: After blocking, the membranes were incubated with the following primary antibodies: Nsun2 (1:1000, PH6626, ab-mart, Shanghai, China), Mettl3 (1:1000, 15,073-1-AP, Proteintech, Rosemont, IL, United States), Ythdf3 (1:500, 25,537-1-AP, Proteintech), FTO (1:1000, PA2776, ab-mart); Wdr4 (1:1000, PS17092, ab-mart), Eif4e (1:1000, 11,149-1-AP, Proteintech), and GAPDH (1:20000, 10,494-1-AP, Proteintech) overnight at 4 °C.

Techniques: Biomarker Discovery, Expressing, Ligation, Western Blot