Journal: Molecular Medicine Reports

Article Title: 7-Difluoromethyl-5,4′-dimethoxygenistein exerts anti-angiogenic effects on acute promyelocytic leukemia HL-60 cells by inhibiting the TLR4/NF-κB signaling pathway

doi: 10.3892/mmr.2020.11029

Figure Lengend Snippet: Effect of DFMG on the TLR4/NF-κB signaling pathway and the protein and mRNA expression of VEGF in HL-60 cells. (A) Protein expression of TLR4, P-NF-κB, IκB-α and VEGF in HL-60 cells. (B) mRNA expression of VEGF and TLR4 in HL-60 cells. There was no statistical difference between the blank control and the solvent control. *P<0.05 vs. solvent control group. DFMG, difluoromethyl-5,4′-dimethoxygenistein; TLR4, toll-like receptor 4; VEGF, vascular endothelial growth factor; P, phosphorylated; NF, nuclear factor.

Article Snippet: The primary antibodies included VEGF (1:10,000; Abcam; cat. no. ab52917), TLR4 (1:1,000; ABclonal Biotech Co., Ltd.; cat. no. A11226), NF-κB p65 (1:1,000; Abbkine Scientific Co., Ltd.; cat. no. Abp57495), phosphorylated (P)-NF-κB p65 (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. AP0123), IκB-α (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. A1187), and β-actin (1:10,000; Abbkine Scientific Co., Ltd.; cat. no. Abp50151).

Techniques: Expressing, Control, Solvent

Journal: Molecular Medicine Reports

Article Title: 7-Difluoromethyl-5,4′-dimethoxygenistein exerts anti-angiogenic effects on acute promyelocytic leukemia HL-60 cells by inhibiting the TLR4/NF-κB signaling pathway

doi: 10.3892/mmr.2020.11029

Figure Lengend Snippet: Effect of toll-like receptor 4 blocker, TAK-242, and activator, LPS, on the protein expression of P-NF-κB and IκB-α in HL-60 cells. *P<0.05 vs. solvent control group. LPS, lipopolysaccharide; P, phosphorylated; NF, nuclear factor.

Article Snippet: The primary antibodies included VEGF (1:10,000; Abcam; cat. no. ab52917), TLR4 (1:1,000; ABclonal Biotech Co., Ltd.; cat. no. A11226), NF-κB p65 (1:1,000; Abbkine Scientific Co., Ltd.; cat. no. Abp57495), phosphorylated (P)-NF-κB p65 (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. AP0123), IκB-α (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. A1187), and β-actin (1:10,000; Abbkine Scientific Co., Ltd.; cat. no. Abp50151).

Techniques: Expressing, Solvent, Control

Journal: Molecular Medicine Reports

Article Title: 7-Difluoromethyl-5,4′-dimethoxygenistein exerts anti-angiogenic effects on acute promyelocytic leukemia HL-60 cells by inhibiting the TLR4/NF-κB signaling pathway

doi: 10.3892/mmr.2020.11029

Figure Lengend Snippet: Protein and mRNA expression of VEGF in HL-60 cells treated with DFMG after activation or inhibition of the TLR4/NF-κB signaling pathway. (A) Protein expression of P-NF-κB, IκB-α and VEGF in HL-60 cells. (B) mRNA expression of VEGF in HL-60 cells. *P<0.05 vs. solvent control group; # P<0.05 vs. DFMG-only group. VEGF, vascular endothelial growth factor; DFMG, difluoromethyl-5,4′-dimethoxygenistein; TLR4, toll-like receptor 4; NF, nuclear factor; P, phosphorylated.

Article Snippet: The primary antibodies included VEGF (1:10,000; Abcam; cat. no. ab52917), TLR4 (1:1,000; ABclonal Biotech Co., Ltd.; cat. no. A11226), NF-κB p65 (1:1,000; Abbkine Scientific Co., Ltd.; cat. no. Abp57495), phosphorylated (P)-NF-κB p65 (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. AP0123), IκB-α (1:2,000; ABclonal Biotech Co., Ltd.; cat. no. A1187), and β-actin (1:10,000; Abbkine Scientific Co., Ltd.; cat. no. Abp50151).

Techniques: Expressing, Activation Assay, Inhibition, Solvent, Control

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: Primers of qRT-PCR

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Sequencing

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: Th17/Treg cell differentiation ratio and TWEAK/Fn14 signaling level in AC mice. ( A ) The state of mice ocular surface was observed by slit-lamp. ( B ) The clinical scores of mice were performed according to the status of eyelid, conjunctiva and cornea. ( C ) HE staining was utilized to evaluate the pathological changes of conjunctival tissue in mice. ( D ) The proportion of Th17 or Treg cells in spleen of mice was assessed by flow cytometry. ( E ) The levels of Th17 and Treg cytokines in mice spleen were evaluated by ELISA. ( F-H ) TWEAK and Fn14 levels in ocular conjunctival tissue were measured by qRT-PCR and WB. ( I ) IHC was used to observe the protein level of TWEAK and Fn14 in ocular conjunctival tissue. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Cell Differentiation, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: TWEAK knockdown affected conjunctivitis in AC mice. ( A ) The effect of TWEAK knockdown on the state of mice ocular surface was observed under slit-lamp. ( B ) Clinical score of eyelid, conjunctiva and cornea to assess the effect of TWEAK knockdown in AC mice. ( C-D ) HE staining and TB staining were utilized to evaluate the effect of TWEAK knockdown on conjunctivitis in mice. ( E-G ) The levels of TWEAK and Fn14 in conjunctival tissue were detected by qRT-PCR, IHC and WB. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Knockdown, Staining, Quantitative RT-PCR

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: TWEAK regulated the Th17/Treg cell ratio in AC mice. ( A ) The effect of TWEAK knockdown on Th17 and Treg cell ratio in spleen of AC mice was observed by flow cytometry. ( B-C ) WB and IHC were employed to evaluate the expression levels of FoxP3 and RORγt in mice conjunctival tissue. ( D ) The effect of TWEAK knockdown on the levels of Th17 and Treg cytokines in mice spleen were evaluated by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Knockdown, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: TWEAK knockdown promoted Nrf2/HO-1 signaling pathway in AC mice. ( A ) qRT-PCR was utilized to measure the effect of TWEAK knockdown on Nrf2 and HO-1 mRNA levels in conjunctival tissue of mice. ( B-C ) The effect of TWEAK knockdown on protein levels of Nrf2 and HO-1 in conjunctival tissue of mice were detected by WB and IHC. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Knockdown, Quantitative RT-PCR, Paraffin-embedded Immunohistochemistry

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: Inhibition of Nrf2/HO-1 signaling pathway affected the improvement of conjunctivitis in AC mice from TWEAK knockdown. ( A ) The effect of Nrf2 inhibitor on ocular surface status was observed by slit-lamp in AC mice with TWEAK knockdown. ( B ) Clinical score of eyelid, conjunctiva and cornea to assess the effect of Nrf2 inhibitor in AC mice with TWEAK knockdown. ( C-D ) HE staining and TB staining were employed to evaluate the effect of Nrf2 inhibitor on conjunctivitis in AC mice with TWEAK knockdown. ( E-G ) The levels of Nrf2 and HO-1 in conjunctival tissue were detected by qRT-PCR, IHC and WB. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. AC + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shTWEAK

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Inhibition, Knockdown, Staining, Quantitative RT-PCR

Journal: Molecular Medicine

Article Title: TWEAK/Fn14 disrupts Th17/Treg balance and aggravates conjunctivitis by inhibiting the Nrf2/HO-1 pathway in allergic conjunctivitis mice

doi: 10.1186/s10020-024-01004-5

Figure Lengend Snippet: Inhibition of Nrf2/HO-1 signaling pathway affected Th17/Treg cell ratio in AC mice with TWEAK knockdown. ( A ) The effect of Nrf2 inhibitor on Th17/Treg cell ratio in AC mice with TWEAK knockdown was assessed by flow cytometry. ( B-C ) WB and IHC assays were employed to evaluate the expression of FoxP3 and RORγt in conjunctival tissue of AC mice. ( D ) The levels of Th17 and Treg cytokines in mice spleen were detected by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. AC + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shTWEAK

Article Snippet: Membranes were blocked by BSA (5%) for 1 h at room temperature and then cultured with primary antibodies against RORγt (ab207082, 1:1000; Abcam), FoxP3 (ab20034, 1:1000; Abcam), TWEAK (BM4635, 1:1000; Boster), Fn14 (A11813, 1:1000; Boster), Nrf2 (ab62352, 1:1000; Abcam), HO-1 (ab68477, 1:1000; Abcam) and GAPDH (ab8245, 1:1000; Abcam) at 4 °C overnight.

Techniques: Inhibition, Knockdown, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction

doi: 10.1016/j.isci.2024.111259

Figure Lengend Snippet:

Article Snippet: Na2HPO4 , Thermo Fisher , Cat# A11817.30.

Techniques: Virus, Recombinant, Protease Inhibitor, Mutagenesis, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: Associations of METTL16 expression with clinical parameters in gastric cancer

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: METTL16 is highly expressed in gastric cancer cell lines and tumour tissues. (A) The mRNA level of METTL16 in normal tissues and GC tissues downloaded from The Cancer Genome Atlas (TCGA) database (** P < .01). (B) mRNA level of METTL16 in normal control and different stages (N0‐N4) of GC patients downloaded from the TCGA database (* P < .05,** P < .01 compared with normal tissues). (C) Western blotting was performed to detect the protein level of METTL16 in 16 pairs of GC tissues and their paired normal adjacent tissues. (D and E) The mRNA level of METTL16 in 16 pairs of GC tissues and their paired normal adjacent tissues (D) and the summarized data (E) (** P < .01). (F) mRNA levels of METTL16 in 6 GC cell lines (AGS, SGC7901, SNU719, MGC803, HGC27, MKN28) and 1 normal gastric mucosal cell line (GES‐1). (G) Protein level of METTL16 in 6 GC cell lines and 1 normal gastric mucosal cell line (GES‐1)

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: High expression of METTL16 in GC is related to poor survival of patients. (A and B) Immunohistochemical staining of METTL16 in GC tissues and paired normal adjacent tissues. (A) Representative image of GC tissues with negative (A and B), weak positive (C and D), moderately positive (E and F) and strong positive (g and h) METTL16 expression. The images are magnified 100X (A, C, E and G) and 400X (B, D, F and H). (B) Representative image of METTL16 staining in GC tissues and the paired normal adjacent tissues (NAT) (magnified 400X). (C) H score of METTL16 staining in GC tissue and the paired normal adjacent tissues ( n = 40, H score 43.63 ± 28.41 vs 79.65 ± 42.80,*** P ﹤ .001 compared with paired normal adjacent tissues).(D) COX regression model for multivariate analysis showed that the expression level of METTL16 was significantly negatively correlated with the overall survival rate in 231 GC patients (log‐ranch test). (E) The expression level of METTL16 in GC patients was significantly negatively correlated with the overall survival rate in the Kaplan‐Meier plotter database (238931_at)

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: Cox proportional hazard regression analysis for overall survival

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: Knock‐down of METTL16 inhibits the proliferation of GC cells in vitro. (A) The mRNA expression of METTL16 in three GC cell lines (AGS, MGC803 and SNU719) after treated with METTL16 shRNA lentivirus. (B) The protein expression of METTL16 in three GC cell lines after treated with METTL16 shRNA lentivirus. (C) Knock‐down of METTL16 can effectively inhibit cell growth in AGS, MGC803 and SNU719 cells. (D and E) Colony formation of AGS, MGC803 and SNU719 cells after transfection of shMETTL16 or shNC. All experiments were performed in triplicate. * P < .05, ** P < .01

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: In Vitro, Expressing, shRNA, Transfection

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: Down‐regulation of METTL16 inhibits the proliferation of GC cells through regulating the cell cycle. (A‐D) Representative immunofluorescence image showing the expression of EdU in AGS (A and B) and MGC803 cells (C and D). (E‐H) Flow cytometry was performed to analyse the cell cycle in shNC‐ or shMETTL16‐treated AGS (E and F) and MGC803 cells (G and H). The knock‐down of METTL16 effectively inhibited the G1/S transition in AGS (E and F) and MGC803 cells (G and H). All results are expressed as the ± SD of three repeated experiments (*0.01 ≤ P < .05; **0.001 ≤ P < .01, *** P < .001)

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: METTL16 promotes tumour growth in vivo. (A) Nude mice are implanted with shNC or shMETTL16‐2 GC cells subcutaneously. (B) Subcutaneous tumour nodules formed in two groups of mice. (C) The growth curve of subcutaneous tumour volume in two groups of mice. (D) Comparison of tumour mass using independent Student's t‐test. (E) HE staining of two groups of tumour specimens. (F) Two groups of tumour specimens were subjected to immunohistochemical detection of METTL16 and Ki67 indicators

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: In Vivo, Staining, Immunohistochemical staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: Knock‐down of METTL16 inhibits the expression of cyclin D1 in GC cells. (A) Western blotting analysis was performed to detect the expression of proteins associated with G1/S phase transition, including cyclin D1, cyclin E1, p21, CDK2 and CDK6. (B) qPCR analysis of the expression of cyclin D1, cyclin E1, p21, p27, CDK2 and CDK6. (C) Representative immunofluorescence image showing the expression of cyclin D1 and METTL16 in subcutaneous tumour tissue of mice

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Expressing, Western Blot, Sublimation, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: METTL16 promotes cell proliferation by up‐regulating cyclin D1 expression in gastric cancer

doi: 10.1111/jcmm.16664

Figure Lengend Snippet: METTL16 regulates cyclin D1 expression via mediating the stability of cyclin D1 mRNA. (A) Based on the standard curve, RNA methylation quantitative analysis method was used to detect the overall content of m6A in shNC‐ or shMETTL16‐treated GC cells. (B) Cyclin D1 expression in shNC‐ or shMETTL16‐treated GC cells induced by actinomycin D for 8 h. (C) The RNA degradation rate was determined in shNC‐ or shMETTL16‐treated GC cells (referring to 0h). (D) The global level of m6A methylation in GC cells treated with different concentrations of DAA (0 umol/L, 100 umol/L, 200 umol/L) for 24 h. (E) The mRNA level of cyclin D1 in AGS, MGC803 and SNU719 cells treated with different concentrations of DAA (0 umol/L, 100 umol/L, 200 umol/L) for 24 h. (F) Western blotting analysis was performed to detect cyclin D1 expression in AGS, MGC803, SNU719 cells treated with different concentrations of DAA (0 umol/L, 100 umol/L, 200 umol/L) for 24 h

Article Snippet: Rabbit anti‐METTL16 polyclonal antibody (1:200; cat. no. A118157; SIGMA) was used overnight at 4°C.

Techniques: Expressing, Methylation, Western Blot