Journal: International Journal of Molecular Sciences

Article Title: PGC1α Cooperates with FOXA1 to Regulate Epithelial Mesenchymal Transition through the TCF4-TWIST1

doi: 10.3390/ijms23158247

Figure Lengend Snippet: Suppression of TCF4 and TWIST1 by ID1 is linked to loss of FOXA1-induced EMT. ( A ) Changes of EMT−associated genes by FOXA1 silencing in ectopic GFP or ID1 expressing A549 cells. ( B ) Interaction between TCF4 and TWIST1 in FOXA1 knockdown A549 cells. ( C ) Promoter occupancy of TCF4 and TWIST1 on CDH1 and CDH2 promoter in FOXA1 silenced A549 cells. ( D ) Changes of EMT-associated genes by FOXA1 silencing in control or TCF4 knockdown A549 cells. Values represent mean ± SD (n = 4). * p < 0.05 and ** p < 0.01 by one-way ANOVA with Tukey post-test for multiple comparisons. Not significate ns is normally this meaning.

Article Snippet: Human TCF4 targeting siRNA (sc-61657) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Knockdown, Control

Journal: International Journal of Molecular Sciences

Article Title: PGC1α Cooperates with FOXA1 to Regulate Epithelial Mesenchymal Transition through the TCF4-TWIST1

doi: 10.3390/ijms23158247

Figure Lengend Snippet: Suppression of TCF4 and TWIST1 by ID1 involves TGFβ1−mediated EMT gene expression. ( A ) Interaction of TCF4 and TWIST1 in the absence or presence of TGFβ1 (20 ng/mL for 24 h). ( B ) EMT-associated gene expression upon TGFβ1 (20 ng/mL for 24 h) in GFP or ID1 stably expressing A549 cells. ( C ) TGFβ1−mediated EMT gene expression in control or TCF4 silenced A549 cells. ( D ) Promoter occupancy of TCF4 on CDH1 and CDH2 promoter in GFP, FOXA1 or PGC1α stably expressing A549 cells. Values represent mean ± SD (n = 3). * p < 0.05 and * * p < 0.01 by one-way ANOVA with Tukey post−test for multiple comparisons. Not significate ns is normally this meaning.

Article Snippet: Human TCF4 targeting siRNA (sc-61657) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Gene Expression, Stable Transfection, Expressing, Control

Journal: Drug Design, Development and Therapy

Article Title: Protective effect of Huang Gan formula in 5/6 nephrectomized rats by depressing the Wnt/β-catenin signaling pathway

doi: 10.2147/DDDT.S81157

Figure Lengend Snippet: Average optical density values of protein expressions by immunohistochemistry staining in renal tissues of rats in different groups, x ¯ + s ( n = 3 )

Article Snippet: Rabbit polyclonal antibody Wnt1 (ab85060; Abcam, Inc., Cambridge, MA, USA), β-catenin (BS1982; Bioworld Technology, Inc., St Louis Park, MN, USA), GSK-3β (sc9166; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-GSK-3β (sc-135653; Santa Cruz Biotechnology Inc., Dallas, TX, USA), transcription factor 4 (TCF4) (BS6172; Bioworld Technology, Inc.), dickkopf 1 (Dkk1) (BS7731; Bioworld Technology, Inc.), fibronectin 1 (Fn1) (ab2413; Abcam, Inc.), and glyceraldehyde 3-phosphate dehydro-genase (GAPDH) (ab9485; Abcam, Inc.) were purchased.

Techniques: Immunohistochemistry, Staining

Journal: Drug Design, Development and Therapy

Article Title: Protective effect of Huang Gan formula in 5/6 nephrectomized rats by depressing the Wnt/β-catenin signaling pathway

doi: 10.2147/DDDT.S81157

Figure Lengend Snippet: Representative micrographs of immunohistochemistry staining in the 5/6 Nx group rat renal cortex sections (400×). Notes: Immunohistochemistry staining of 5/6 Nx group for Wnt1 ( A ), β-catenin ( B ), GSK-3β ( C ), Dkk1 ( D ), TCF4 ( E ), Fn1 ( F ), and IgG negative control ( G ). Abbreviations: Nx, 5/6 nephrectomy; GSK-3β, glycogen synthase kinase-3β; Dkk1, dickkopf 1; TCF4, transcription factor 4; Fn1, fibronectin 1.

Article Snippet: Rabbit polyclonal antibody Wnt1 (ab85060; Abcam, Inc., Cambridge, MA, USA), β-catenin (BS1982; Bioworld Technology, Inc., St Louis Park, MN, USA), GSK-3β (sc9166; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-GSK-3β (sc-135653; Santa Cruz Biotechnology Inc., Dallas, TX, USA), transcription factor 4 (TCF4) (BS6172; Bioworld Technology, Inc.), dickkopf 1 (Dkk1) (BS7731; Bioworld Technology, Inc.), fibronectin 1 (Fn1) (ab2413; Abcam, Inc.), and glyceraldehyde 3-phosphate dehydro-genase (GAPDH) (ab9485; Abcam, Inc.) were purchased.

Techniques: Immunohistochemistry, Staining, Negative Control

Journal: Drug Design, Development and Therapy

Article Title: Protective effect of Huang Gan formula in 5/6 nephrectomized rats by depressing the Wnt/β-catenin signaling pathway

doi: 10.2147/DDDT.S81157

Figure Lengend Snippet: Wnt1, β-catenin, TCF4, GSK-3β, Dkk1, and Fn1 mRNA expression in renal tissues (n=6). Notes: Compared with the 5/6 Nx group, * P <0.05; compared with Sham and BC groups, # P <0.05; and compared with the losartan group, Δ P <0.05. Abbreviations: TCF4, transcription factor 4; GSK-3β, glycogen synthase kinase-3β; Dkk1, dickkopf 1; Fn1, fibronectin 1; Nx, 5/6 nephrectomy; Sham, sham-operated group; BC, background control group; UCG, uremic clearance granule; HGF-L, Huang Gan formula low dose group; HGF-M, Huang Gan formula middle dose group; HGF-H, Huang Gan formula high dose group.

Article Snippet: Rabbit polyclonal antibody Wnt1 (ab85060; Abcam, Inc., Cambridge, MA, USA), β-catenin (BS1982; Bioworld Technology, Inc., St Louis Park, MN, USA), GSK-3β (sc9166; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-GSK-3β (sc-135653; Santa Cruz Biotechnology Inc., Dallas, TX, USA), transcription factor 4 (TCF4) (BS6172; Bioworld Technology, Inc.), dickkopf 1 (Dkk1) (BS7731; Bioworld Technology, Inc.), fibronectin 1 (Fn1) (ab2413; Abcam, Inc.), and glyceraldehyde 3-phosphate dehydro-genase (GAPDH) (ab9485; Abcam, Inc.) were purchased.

Techniques: Expressing

Journal: Drug Design, Development and Therapy

Article Title: Protective effect of Huang Gan formula in 5/6 nephrectomized rats by depressing the Wnt/β-catenin signaling pathway

doi: 10.2147/DDDT.S81157

Figure Lengend Snippet: Western blot analysis for detection of Wnt1, β-catenin, TCF4, GSK-3β, Dkk1, and Fn1 expression in the kidney. Notes: Western blot bands of different proteins ( A ); semi-quantitative analysis of the bands (from ( B ) to ( G ) are Wnt1, β-catenin, GSK-3β, Dkk1, TCF4, and Fn1 in sequence). Compared with 5/6 Nx group, * P <0.05; compared with Sham and BC groups, # P <0.05; and compared with the losartan group Δ P <0.05. Abbreviations: TCF4, transcription factor 4; GSK-3β, glycogen synthase kinase-3β; Dkk1, dickkopf 1; Fn1, fibronectin 1; Nx, 5/6 nephrectomy; Sham, sham-operated group; BC, background control group; UCG, uremic clearance granule; HGF-L, Huang Gan formula low dose group; HGF-M, Huang Gan formula middle dose group; HGF-H, Huang Gan formula high dose group; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Rabbit polyclonal antibody Wnt1 (ab85060; Abcam, Inc., Cambridge, MA, USA), β-catenin (BS1982; Bioworld Technology, Inc., St Louis Park, MN, USA), GSK-3β (sc9166; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-GSK-3β (sc-135653; Santa Cruz Biotechnology Inc., Dallas, TX, USA), transcription factor 4 (TCF4) (BS6172; Bioworld Technology, Inc.), dickkopf 1 (Dkk1) (BS7731; Bioworld Technology, Inc.), fibronectin 1 (Fn1) (ab2413; Abcam, Inc.), and glyceraldehyde 3-phosphate dehydro-genase (GAPDH) (ab9485; Abcam, Inc.) were purchased.

Techniques: Western Blot, Expressing, Sequencing

Journal: Drug Design, Development and Therapy

Article Title: Protective effect of Huang Gan formula in 5/6 nephrectomized rats by depressing the Wnt/β-catenin signaling pathway

doi: 10.2147/DDDT.S81157

Figure Lengend Snippet: Primers used for RT-PCR assay

Article Snippet: Rabbit polyclonal antibody Wnt1 (ab85060; Abcam, Inc., Cambridge, MA, USA), β-catenin (BS1982; Bioworld Technology, Inc., St Louis Park, MN, USA), GSK-3β (sc9166; Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-GSK-3β (sc-135653; Santa Cruz Biotechnology Inc., Dallas, TX, USA), transcription factor 4 (TCF4) (BS6172; Bioworld Technology, Inc.), dickkopf 1 (Dkk1) (BS7731; Bioworld Technology, Inc.), fibronectin 1 (Fn1) (ab2413; Abcam, Inc.), and glyceraldehyde 3-phosphate dehydro-genase (GAPDH) (ab9485; Abcam, Inc.) were purchased.

Techniques: Sequencing, Amplification

Journal: Cell death & disease

Article Title: Aberrant Gcm1 expression mediates Wnt/β-catenin pathway activation in folate deficiency involved in neural tube defects.

doi: 10.1038/s41419-020-03313-z

Figure Lengend Snippet: Fig. 1 Wnt/β-catenin pathway signaling activation in cells with folate deficiency. a Morphology of C57BL/6 mESCs as observed with an optical microscope at magnifications of ×100 and ×200, F6, sixth generation of C57BL/6 mESCs. b Analysis of the cell cycle distribution of C57BL/6 mESCs after six generations of culture under folate deficiency by flow cytometry. 3 × 106 cells were harvested for control and case group respectively; right panel: the results of cell cycle quantification were graded according to the cell cycle phase. c Analysis of apoptosis in C57BL/6 mESCs after six generations of culture under folate deficiency by flow cytometry. 3 × 106 cells were harvested for control and case group, respectively, the number of cells in early apoptosis is shown in a bar graph in the right panel. d Folate concentration in C57BL/6 mESCs after six generations of culture in folate deficiency and in NE-4C after three generations of culture in folate deficiency. e The mRNA expression of Fzd5, Lrp, Wnt6, Ccnd1, Nfat5, Lef1, TCF4, TCF12, Bcl9l, and Axin2 in C57BL/6 mESCs with folate deficiency. f Analysis of luciferase activity by TOP/FOP Flash assays in C57BL/6 mESCs subjected to folate deficiency for six generations and NE-4C subjected to folate deficiency for three generations. Data a–f represent the mean ± SEM (n = 3). The p value was calculated by Student’s t-test, ns was for no significance, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: siRNA reagents for Gcm1 and TCF4 were purchased from Santa Cruz Biotechnology (USA).

Techniques: Activation Assay, Microscopy, Cytometry, Control, Concentration Assay, Expressing, Luciferase, Activity Assay

Journal: The Journal of Clinical Investigation

Article Title: FOXOs attenuate bone formation by suppressing Wnt signaling

doi: 10.1172/JCI68049

Figure Lengend Snippet: (A) Wnt-target genes determined by microarray analysis in Osx1-GFP calvaria cells. (B) mRNA levels by qRT-PCR in calvaria cells treated with vehicle (veh) or 50 ng/ml Wnt3a overnight (triplicates). (C) mRNA levels by qRT-PCR in femoral shafts from 3-month-old mice (n = 8–9/group), and in periosteal and bone marrow osteoblastic cell cultures described in Figure ​Figure4,4, C and G. (D) mRNA by qRT-PCR (left, triplicates) and protein levels by Western blot (right) in calvaria-derived cells infected with Adeno-Cre to induce Foxo deletion. (E) BrdU incorporation in cells, described in D, treated with veh (–) or 50 ng/ml Wnt3a (triplicates). (F) Lysates from nuclear extracts of calvaria cells, described in D, treated with vehicle or 50 ng/ml Wnt3a, immunoprecipitated with an anti–TCF-4 or anti-IgG antibody, and probed with an anti–β-catenin antibody. (G) mRNA levels by qRT-PCR and (H) BrdU incorporation in calvaria cells infected with Adeno-Cre and with lentivirus expressing shRNA directed against β-catenin or nontargeted shRNA followed by treatment with vehicle or 50 ng/ml Wnt3a overnight (triplicates). #P < 0.05 by Student’s t test; *P < 0.05 by ANOVA with Bonferroni’s test. Bars represent mean + SD.

Article Snippet: To analyze β-catenin–TCF-4 interactions, cell lysates were immunoprecipitated with an anti–TCF-4 antibody (#2569S; Cell Signaling) or an IgG nonspecific control antibody (sc-2003; Santa Cruz Biotechnology Inc.).

Techniques: Microarray, Quantitative RT-PCR, Western Blot, Derivative Assay, Infection, BrdU Incorporation Assay, Immunoprecipitation, Expressing, shRNA