Journal: International Journal of Molecular Sciences

Article Title: The Antitumor Effect of Timosaponin A3 through c-Myc Inhibition in Colorectal Cancer Cells and Combined Treatment Effect with 5-FU or Doxorubicin

doi: 10.3390/ijms231911900

Figure Lengend Snippet: TA3 inhibits c-Myc and induces apoptosis via CNOT2 or MID1IP1. ( A ) HCT116 p53+/+ cells were transfected with CNOT2 siRNA or control siRNA for 48 h and then cells were treated with TA3 (12.5 μM) for 24 h (three independent experiments in HCT116 p53+/+ cells). ( B ) HCT116 p53+/+ cells were transfected with MID1IP1 siRNA or control siRNA for 48 h and then cells were treated with TA3 for 24 h (three independent experiments in HCT116 p53+/+ cells). Statistical significance was analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s test and the unpaired t-test. p -value indicates a significant difference between the groups. Quantification of protein expression is shown in the lower panel. * p < 0.01, ** p < 0.005, *** p < 0.001 vs. control; # p < 0.01, ## p < 0.005, ### p < 0.001 vs. only TA3-treated group. ( C ) HCT116 p53+/+ cells were treated with TA3 for 24 h. Then, cells were harvested for co-immunoprecipitation assay.

Article Snippet: Primary antibodies for CNOT2 (Cat No. 34214) and PARP (Cat No. 9542) were purchased from Cell Signaling Technology (Beverly, MA, USA), MID1IP1 (Cat No.15764-1) was purchased from Proteintech (Rosemont, IL, USA), and c-Myc(Y69) (Cat No. 32072) was purchased from Abcam (Cambridge, UK).

Techniques: Transfection, Control, Expressing, Co-Immunoprecipitation Assay

Journal: International Journal of Molecular Sciences

Article Title: The Antitumor Effect of Timosaponin A3 through c-Myc Inhibition in Colorectal Cancer Cells and Combined Treatment Effect with 5-FU or Doxorubicin

doi: 10.3390/ijms231911900

Figure Lengend Snippet: Mechanism of TA3 to induce apoptosis in colorectal cancer by downregulating c-Myc via CNOT2 or MID1IP1.

Article Snippet: Primary antibodies for CNOT2 (Cat No. 34214) and PARP (Cat No. 9542) were purchased from Cell Signaling Technology (Beverly, MA, USA), MID1IP1 (Cat No.15764-1) was purchased from Proteintech (Rosemont, IL, USA), and c-Myc(Y69) (Cat No. 32072) was purchased from Abcam (Cambridge, UK).

Techniques:

Journal: Scientific Reports

Article Title: G protein-coupled receptor 91 activations suppressed mineralization in Porphyromonas gingivalis –infected osteoblasts

doi: 10.1038/s41598-024-78944-9

Figure Lengend Snippet: P. gingivalis infection promoted GPR91 expression with inhibited mineralization in osteoblasts. Osteoblasts were cultured with different multiplicity of infections (MOIs) of P. gingivalis , and the expressions of OSX, RUNX2, OPN and IL-6 were detected by real-time PCR for 24 h ( A ) and western blotting after 48 h stimulation ( B ). ALP staining ( C ) was performed at 7 days, and ARS ( D ) was performed at 14 days after being stimulated with P. gingivalis at a MOI of 50. Expressions of mineralization-related markers and GPR91 in osteoblasts cementogenic-differentiated at 0, 4, and 7 days were examined by qPCR ( E ) and western blotting ( F ). In all cases, bars in graphs represent mean ± SEM. β-actin was adopted as an internal reference. *, p < 0.05; **, p < 0.01 compared with the Control.

Article Snippet: The membrane was blocked with 5% bovine albumin and then incubated with primary antibodies: OSX (1:1000; A18699, ABclonal, China), RUNX2 (1:1000; D1L7F, CST, Germany), OPN (1:1000; A21084, ABclonal, China), GPR91 (1:1000, orb157370, Biorbyt, China), RANKL (1:1000, 23408-1-AP, PTG, China), OPG (1:1000, DF6824, Affinity, China), TRAP (1:1000; A0962, ABclonal, China), MMP9 (1:1000; A11147, ABclonal, China), CCL2 (1:1000; A23288, ABclonal, China), P38 (1:1000; 8690, CST, Germany), p-P38 (1:1000; 4511, CST, Germany), JNK (1:1000; 9252, CST, Germany), p-JNK (1:1000; 4668, CST, Germany), p-P65 (1:1000, 93H1, CST, Germany), ERK (1:1000, GB11560, Servicebio, China), p-ERK (1:1000, AF1015, Affinity, China), β-actin (1:1000; 66009-l-lg, Proteintech, China ).

Techniques: Infection, Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control

Journal: Scientific Reports

Article Title: G protein-coupled receptor 91 activations suppressed mineralization in Porphyromonas gingivalis –infected osteoblasts

doi: 10.1038/s41598-024-78944-9

Figure Lengend Snippet: Blocking GPR91 mitigated the bone mineralization inhibited by P.gingivalis. CCK8 examined the activity of osteoblasts treated with 4C at different concentrations ( A ). Osteoblasts were pretreated with 4C (5 μM) for 2 h and then treated with P. gingivalis at a MOI of 50. Gene transcript levels of OSX, RUNX2, OPN and IL-6 were analyzed by real-time PCR at 24 h ( B ) and protein levels were detected by western blotting after 48 h stimulation ( C ). ALP staining and ALP activity assay at 7 days ( D ) and ARS at 14 days ( E ) of osteoblasts treated with P. gingivalis at a MOI of 50. In all cases, bars in graphs represent mean ± SEM. β-actin was adopted as an internal reference. *, p < 0.05; **, p < 0.01 compared with the Control; #, p < 0.05; ##, p < 0.01 compared with the P. gingivalis -treated group.

Article Snippet: The membrane was blocked with 5% bovine albumin and then incubated with primary antibodies: OSX (1:1000; A18699, ABclonal, China), RUNX2 (1:1000; D1L7F, CST, Germany), OPN (1:1000; A21084, ABclonal, China), GPR91 (1:1000, orb157370, Biorbyt, China), RANKL (1:1000, 23408-1-AP, PTG, China), OPG (1:1000, DF6824, Affinity, China), TRAP (1:1000; A0962, ABclonal, China), MMP9 (1:1000; A11147, ABclonal, China), CCL2 (1:1000; A23288, ABclonal, China), P38 (1:1000; 8690, CST, Germany), p-P38 (1:1000; 4511, CST, Germany), JNK (1:1000; 9252, CST, Germany), p-JNK (1:1000; 4668, CST, Germany), p-P65 (1:1000, 93H1, CST, Germany), ERK (1:1000, GB11560, Servicebio, China), p-ERK (1:1000, AF1015, Affinity, China), β-actin (1:1000; 66009-l-lg, Proteintech, China ).

Techniques: Blocking Assay, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Staining, ALP Activity Assay, Control

Journal: Scientific Reports

Article Title: G protein-coupled receptor 91 activations suppressed mineralization in Porphyromonas gingivalis –infected osteoblasts

doi: 10.1038/s41598-024-78944-9

Figure Lengend Snippet: GPR91 knockdown mitigated the bone mineralization inhibited by P.gingivalis . Osteoblasts from WT and GPR91 -/- mice were stimulated with P. gingivalis (MOI = 50). Gene transcript levels of OSX, RUNX2, OPN and IL-6 were analyzed by real-time PCR for 24 h ( A ), and protein levels were detected by western blotting after 48 h stimulation ( B ). ALP staining and ALP activity assay at 7 days ( C ) and ARS at 14 days ( D ) of osteoblasts treated with P. gingivalis at a MOI of 50. In all cases, bars in graphs represent mean ± SEM. β-actin was adopted as an internal reference. *, p < 0.05; **, p < 0.01 compared with the WT group; #, p < 0.05; ##, p < 0.01 compared with the WT + P. g -treated group. The WT group served as the Control group.

Article Snippet: The membrane was blocked with 5% bovine albumin and then incubated with primary antibodies: OSX (1:1000; A18699, ABclonal, China), RUNX2 (1:1000; D1L7F, CST, Germany), OPN (1:1000; A21084, ABclonal, China), GPR91 (1:1000, orb157370, Biorbyt, China), RANKL (1:1000, 23408-1-AP, PTG, China), OPG (1:1000, DF6824, Affinity, China), TRAP (1:1000; A0962, ABclonal, China), MMP9 (1:1000; A11147, ABclonal, China), CCL2 (1:1000; A23288, ABclonal, China), P38 (1:1000; 8690, CST, Germany), p-P38 (1:1000; 4511, CST, Germany), JNK (1:1000; 9252, CST, Germany), p-JNK (1:1000; 4668, CST, Germany), p-P65 (1:1000, 93H1, CST, Germany), ERK (1:1000, GB11560, Servicebio, China), p-ERK (1:1000, AF1015, Affinity, China), β-actin (1:1000; 66009-l-lg, Proteintech, China ).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Staining, ALP Activity Assay, Control

Journal: Scientific Reports

Article Title: G protein-coupled receptor 91 activations suppressed mineralization in Porphyromonas gingivalis –infected osteoblasts

doi: 10.1038/s41598-024-78944-9

Figure Lengend Snippet: GPR91-NFκB signalling pathway was involved in the mineralization of osteoblasts under inflammation. ( A ) Osteoblasts from WT mice were treated with P. gingivalis (MOI = 50) for 1 h and harvested for western blotting to reveal the phosphorylation of NF-κB and MAPK pathways. Osteoblasts were pretreated with SCH772984 (ERK inhibitor, 500 nM) and SC75741 (P65 inhibitor, 5 μM) and then treated with P. gingivalis (MOI = 50) for 48 h. Protein levels of OSX, RUNX2, OPN and GPR91 were detected by western blotting ( B & C ). In all cases, bars in graphs represent mean ± SEM. *, p < 0.05; **, p < 0.01 compared with the Control group; #, p < 0.05; ##, p < 0.01 compared with the P. g -treated group.

Article Snippet: The membrane was blocked with 5% bovine albumin and then incubated with primary antibodies: OSX (1:1000; A18699, ABclonal, China), RUNX2 (1:1000; D1L7F, CST, Germany), OPN (1:1000; A21084, ABclonal, China), GPR91 (1:1000, orb157370, Biorbyt, China), RANKL (1:1000, 23408-1-AP, PTG, China), OPG (1:1000, DF6824, Affinity, China), TRAP (1:1000; A0962, ABclonal, China), MMP9 (1:1000; A11147, ABclonal, China), CCL2 (1:1000; A23288, ABclonal, China), P38 (1:1000; 8690, CST, Germany), p-P38 (1:1000; 4511, CST, Germany), JNK (1:1000; 9252, CST, Germany), p-JNK (1:1000; 4668, CST, Germany), p-P65 (1:1000, 93H1, CST, Germany), ERK (1:1000, GB11560, Servicebio, China), p-ERK (1:1000, AF1015, Affinity, China), β-actin (1:1000; 66009-l-lg, Proteintech, China ).

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Scientific Reports

Article Title: G protein-coupled receptor 91 activations suppressed mineralization in Porphyromonas gingivalis –infected osteoblasts

doi: 10.1038/s41598-024-78944-9

Figure Lengend Snippet: The primer sequences used for real-time qPCR.

Article Snippet: The membrane was blocked with 5% bovine albumin and then incubated with primary antibodies: OSX (1:1000; A18699, ABclonal, China), RUNX2 (1:1000; D1L7F, CST, Germany), OPN (1:1000; A21084, ABclonal, China), GPR91 (1:1000, orb157370, Biorbyt, China), RANKL (1:1000, 23408-1-AP, PTG, China), OPG (1:1000, DF6824, Affinity, China), TRAP (1:1000; A0962, ABclonal, China), MMP9 (1:1000; A11147, ABclonal, China), CCL2 (1:1000; A23288, ABclonal, China), P38 (1:1000; 8690, CST, Germany), p-P38 (1:1000; 4511, CST, Germany), JNK (1:1000; 9252, CST, Germany), p-JNK (1:1000; 4668, CST, Germany), p-P65 (1:1000, 93H1, CST, Germany), ERK (1:1000, GB11560, Servicebio, China), p-ERK (1:1000, AF1015, Affinity, China), β-actin (1:1000; 66009-l-lg, Proteintech, China ).

Techniques: Sequencing

Journal: Physiological Genomics

Article Title: Human gene copy number spectra analysis in congenital heart malformations

doi: 10.1152/physiolgenomics.00013.2012

Figure Lengend Snippet: Known CHD risk genes

Article Snippet: MID1 , MIDLINE 1 , Xp22.2 , 10373595 , 388135 , Hs02158662_cn , S , 300552 , 12833403, 20193066.

Techniques: Retroviral, Sequencing, Virus, Reverse Transcription

Journal: Physiological Genomics

Article Title: Human gene copy number spectra analysis in congenital heart malformations

doi: 10.1152/physiolgenomics.00013.2012

Figure Lengend Snippet: Case reports of likely causal CNVs

Article Snippet: MID1 , MIDLINE 1 , Xp22.2 , 10373595 , 388135 , Hs02158662_cn , S , 300552 , 12833403, 20193066.

Techniques:

Journal: Physiological Genomics

Article Title: Human gene copy number spectra analysis in congenital heart malformations

doi: 10.1152/physiolgenomics.00013.2012

Figure Lengend Snippet: CHD-associated gene regions significantly enriched with large, rare CNVs

Article Snippet: MID1 , MIDLINE 1 , Xp22.2 , 10373595 , 388135 , Hs02158662_cn , S , 300552 , 12833403, 20193066.

Techniques:

Journal: Physiological Genomics

Article Title: Human gene copy number spectra analysis in congenital heart malformations

doi: 10.1152/physiolgenomics.00013.2012

Figure Lengend Snippet: Enriched syndrome genes

Article Snippet: MID1 , MIDLINE 1 , Xp22.2 , 10373595 , 388135 , Hs02158662_cn , S , 300552 , 12833403, 20193066.

Techniques: