Journal: International Journal of Molecular Sciences

Article Title: NACA and LRP6 Are Part of a Common Genetic Pathway Necessary for Full Anabolic Response to Intermittent PTH

doi: 10.3390/ijms23020940

Figure Lengend Snippet: Naca and Lrp6 form part of a common genetic pathway regulating bone mass. Femurs of female mice were analyzed by μCT at 3 months of age. BV/TV, bone volume/tissue volume. * p < 0.05, two-way ANOVA with post hoc tests.

Article Snippet: Real-time qPCR using TaqMan universal PCR master mix and gene-specific Taqman assays for Lrp6 (Mm00521783_m1), alkaline phosphatase ( Alpl , Mm00475834_m1), and the alpha 1 chain of type I collagen ( Col1a1 , Mm00801666_g1) was performed using a QuantStudio 7 real-time PCR system (Life Technologies Applied Biosystems).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: NACA and LRP6 Are Part of a Common Genetic Pathway Necessary for Full Anabolic Response to Intermittent PTH

doi: 10.3390/ijms23020940

Figure Lengend Snippet: The osteoanabolic response to iPTH is inhibited in Naca / Lrp6 compound heterozygotes. Four-month-old female mice were treated once daily, 5 days/week for 1 month with 100 μg/kg of PTH(1-34). Vertebrae were analyzed by μCT. BV/TV, bone volume/tissue volume. Results are expressed as change from vehicle-treated animals, which are ascribed a value of 100%. * p < 0.05, two-way ANOVA with post hoc tests.

Article Snippet: Real-time qPCR using TaqMan universal PCR master mix and gene-specific Taqman assays for Lrp6 (Mm00521783_m1), alkaline phosphatase ( Alpl , Mm00475834_m1), and the alpha 1 chain of type I collagen ( Col1a1 , Mm00801666_g1) was performed using a QuantStudio 7 real-time PCR system (Life Technologies Applied Biosystems).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: NACA and LRP6 Are Part of a Common Genetic Pathway Necessary for Full Anabolic Response to Intermittent PTH

doi: 10.3390/ijms23020940

Figure Lengend Snippet: The increase in biomechanical properties induced by iPTH treatment is blunted in Naca / Lrp6 compound heterozygotes. Four-month-old female mice were treated once daily, 5 days/week for 1 month with 100 μg/kg of PTH(1-34). L2 vertebrae were analyzed by compression testing to measure stiffness ( A ), load at yield ( B ) and maximum load ( C ). Results are expressed as change from vehicle-treated animals, which are ascribed a value of 100%. * p < 0.05; ** p < 0.01; # p < 0.0001, two-way analysis of variance (ANOVA) with post hoc tests.

Article Snippet: Real-time qPCR using TaqMan universal PCR master mix and gene-specific Taqman assays for Lrp6 (Mm00521783_m1), alkaline phosphatase ( Alpl , Mm00475834_m1), and the alpha 1 chain of type I collagen ( Col1a1 , Mm00801666_g1) was performed using a QuantStudio 7 real-time PCR system (Life Technologies Applied Biosystems).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: NACA and LRP6 Are Part of a Common Genetic Pathway Necessary for Full Anabolic Response to Intermittent PTH

doi: 10.3390/ijms23020940

Figure Lengend Snippet: Gene expression monitoring in control ( Naca 99S/S ; Lrp6 +/+ ;OCN-Cre) and compound heterozygous ( Naca 99S/A ; Lrp6 +/fl ;OCN-Cre) littermates. RNA was extracted from marrow-flushed tibial diaphyses, reverse-transcribed and analyzed by RT-qPCR. Relative expression of Lrp6 ( A ), Alpl ( B ) and Col1a1 ( C ) is shown. * p < 0.05; ** p < 0.01; *** p < 0.001, two-way ANOVA with post hoc tests.

Article Snippet: Real-time qPCR using TaqMan universal PCR master mix and gene-specific Taqman assays for Lrp6 (Mm00521783_m1), alkaline phosphatase ( Alpl , Mm00475834_m1), and the alpha 1 chain of type I collagen ( Col1a1 , Mm00801666_g1) was performed using a QuantStudio 7 real-time PCR system (Life Technologies Applied Biosystems).

Techniques: Gene Expression, Control, Reverse Transcription, Quantitative RT-PCR, Expressing

Journal: International Journal of Molecular Sciences

Article Title: NACA and LRP6 Are Part of a Common Genetic Pathway Necessary for Full Anabolic Response to Intermittent PTH

doi: 10.3390/ijms23020940

Figure Lengend Snippet: Parallel PTH signaling pathways in control ( Naca 99S/S ; Lrp6 +/+ ;OCN-Cre) and compound heterozygous ( Naca 99S/A ; Lrp6 +/fl ;OCN-Cre) littermates. Protein extracts were prepared from tibial shafts and immunoprobed for β-Catenin ( A ) and SIK2 ( B ). Equivalent loading was assessed by probing for α-Tubulin ( C ).

Article Snippet: Real-time qPCR using TaqMan universal PCR master mix and gene-specific Taqman assays for Lrp6 (Mm00521783_m1), alkaline phosphatase ( Alpl , Mm00475834_m1), and the alpha 1 chain of type I collagen ( Col1a1 , Mm00801666_g1) was performed using a QuantStudio 7 real-time PCR system (Life Technologies Applied Biosystems).

Techniques: Protein-Protein interactions, Control

Journal: Translational Vision Science & Technology

Article Title: SZN-413, a FZD4 Agonist, as a Potential Novel Therapeutic for the Treatment of Diabetic Retinopathy

doi: 10.1167/tvst.11.9.19

Figure Lengend Snippet: FZD4 and LRP5, primary receptors for Wnt signaling in vascular endothelial cells. (A) FZD1–10 and LRP5 and LRP6 mRNA expression in HRMECs, bEnd.3 cells, and HUVECs quantified by quantitative polymerase chain reaction (qPCR) and normalized by the housekeeping gene Actin . Bars represent mean ± SD. (B) Data from Macosko et al. 31 were reanalyzed to derive scaled average gene expression by cell type of Wnt receptors in mouse retina. (C) Data from Menon et al. were reanalyzed to derive scaled average gene expression by cell type of Wnt receptors in human retina.

Article Snippet: RNA was quantified using TaqMan Fast Advanced Master Mix (Thermo Fisher Scientific) with the following Thermo Fisher Scientific probes: Mm00445405_s1 Fzd1 , Mm02524776_s1 Fzd2 , Mm00445423_m1 Fzd3 , Mm00433382_m1 Fzd4 , Mm00445623_s1 Fzd5 , Mm00433387_m1 Fzd6 , Mm00433409_s1 Fzd7 , Mm01234717_s1 Fzd8 , Mm01206511_s1 Fzd9 , Mm00558396_s1 Fzd10 , Mm01227476_m1 Lrp5 , Mm00999795_m1 Lrp6 , Hs00268943_s1 FZD1 , Hs00361432_s1 FZD2 , Hs00907280_m1 FZD3 , Hs00201853_m1 FZD4 , Hs00258278_s1 FZD5 , Hs01095627_m1 FZD6 , Hs00275833_s1 FZD7 , Hs00259040_s1 FZD8 , Hs00268954_s1 FZD9 , Hs00273077_s1 FZD10 , Hs00182031_m1 LRP5 , Hs00233945_m1 LRP6 , Hs01547250_m1 LEF1 , Hs00293017_m1 MFSD2A , Hs00610344-m1 AXIN2 , and Mm00443610_m1 Axin2 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Life

Article Title: The Relationship between LRP6 and Wnt/β-Catenin Pathway in Colorectal and Esophageal Cancer

doi: 10.3390/life13030615

Figure Lengend Snippet: LRP6 expression in normal colon and CRC tissues. ( A ) LRP6 expression in normal colon mucosa. ( B , C ) Classification of the positivity of LRP6 expression in CRC. ( D , E ) Classification of LRP6 expression intensity in CRC. ( F ) Distribution of the staining score, calculated by multiplying the intensity and positivity scores.

Article Snippet: The product numbers of the TaqMan Gene Expression assay were as follows: ACTB ID Hs01060665_g1 and LRP6 ID Hs00999795_m1.

Techniques: Expressing, Staining

Journal: Life

Article Title: The Relationship between LRP6 and Wnt/β-Catenin Pathway in Colorectal and Esophageal Cancer

doi: 10.3390/life13030615

Figure Lengend Snippet: LRP6 and β-catenin expression in normal esophageal and ESCC tissues. ( A ) LRP6 expression in normal esophageal tissues. ( B , C ) Distribution of the LRP6 staining score, calculated by multiplying the intensity and positivity scores. ( D , E ) Distribution of the β-catenin staining score, calculated by multiplying the intensity and positivity scores.

Article Snippet: The product numbers of the TaqMan Gene Expression assay were as follows: ACTB ID Hs01060665_g1 and LRP6 ID Hs00999795_m1.

Techniques: Expressing, Staining

Journal: Life

Article Title: The Relationship between LRP6 and Wnt/β-Catenin Pathway in Colorectal and Esophageal Cancer

doi: 10.3390/life13030615

Figure Lengend Snippet: Effects of LRP6 on Wnt/β-catenin signaling activity. ( A ) Expression of LRP6 mRNA in CRC and ESCC cell lines, as measured by qRT-PCR. We used ACTB as endogenous control. * p < 0.01. ( B ) The protein level of LRP6 in CRC and ESCC cell lines. We used ACTB as endogenous control. ( C ) Wnt/β-catenin signaling activity under unstimulated conditions in CRC and ESCC cell lines. ( D ) Experimental plan for investigating signaling activity by adding LRP6-siRNA or Wnt3a to the CRC and ESCC cell lines. ( E – H ) Wnt/β-catenin signaling activity using CRC and ESCC cell lines. We used Renilla as endogenous control. * p < 0.05, ** p < 0.01; n.s.: not significant.

Article Snippet: The product numbers of the TaqMan Gene Expression assay were as follows: ACTB ID Hs01060665_g1 and LRP6 ID Hs00999795_m1.

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Control

Journal: Life

Article Title: The Relationship between LRP6 and Wnt/β-Catenin Pathway in Colorectal and Esophageal Cancer

doi: 10.3390/life13030615

Figure Lengend Snippet: Comparison of clinicopathological status stratified by LRP6 or β-catenin expression in CRC tissue samples.

Article Snippet: The product numbers of the TaqMan Gene Expression assay were as follows: ACTB ID Hs01060665_g1 and LRP6 ID Hs00999795_m1.

Techniques: Comparison, Expressing

Journal: Life

Article Title: The Relationship between LRP6 and Wnt/β-Catenin Pathway in Colorectal and Esophageal Cancer

doi: 10.3390/life13030615

Figure Lengend Snippet: Comparison of clinicopathological status stratified by LRP6 or β-catenin expression in ESCC tissue samples.

Article Snippet: The product numbers of the TaqMan Gene Expression assay were as follows: ACTB ID Hs01060665_g1 and LRP6 ID Hs00999795_m1.

Techniques: Comparison, Expressing

Journal: PLoS ONE

Article Title: Effects of pharmacologic sclerostin inhibition or testosterone administration on soleus muscle atrophy in rodents after spinal cord injury

doi: 10.1371/journal.pone.0194440

Figure Lengend Snippet: A-C. LDL receptor related protein 5 (LRP5) and 6 (LRP6), and androgen receptor (AR) gene expressions in tibia, levator ani/bulbocavernosus (LABC), and soleus from animals receiving sham surgery (T 8 laminectomy). Values are means ± SEM [corrected for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the housekeeping gene] and expressed relative to the tibia, n = 6/tissue. Letters a-c indicate differences from respectively labeled groups at p<0.05 or *p<0.01 (a = vs Tibia; b = vs LABC; c = vs Soleus).

Article Snippet: The following commercially-available gene-specific primers were used (ThermoFisher Scientific, Carlsbad, CA): rat LRP5 (#Rn01451428_m1), rat LRP6 (#Rn01492711_m1), rat AR (#Rn00560747_m1), and rat GAPDH (#Rn01775763_m1).

Techniques: Labeling

Journal: Cancer science

Article Title: Odd-skipped related 1 inhibits lung cancer proliferation and invasion by reducing Wnt signaling through the suppression of SOX9 and β-catenin.

doi: 10.1111/cas.13614

Figure Lengend Snippet: FIGURE 6 Regulation of SOX9 and b-catenin by OSR1. A, The mRNA levels of OSR1, SOX9 and b-catenin (CTNNB1) in H1299-OSR1 cells, cells transfected with pCMV6-SOX9 (H1299-SOX9), cells transfected with both pCMV6-OSR1 and pCMV6-SOX9 (H1299-OSR1 + SOX9) and NC cells. B, The mRNA levels of OSR1, SOX9 and b-catenin (CTNNB1) in A549-siOSR1 cells, cells transfected with SOX9-siRNA (A549-siSOX9), cells transfected with both OSR1-siRNA and SOX9-siRNA (A549-siOSR1 + siSOX9), and siNC cells. C, The western blot analysis of OSR1, SOX9, GSK3b and active b-catenin in H1299-OSR1 cells, H1299-SOX9 cells, H1299-OSR1 + SOX9 cells and NC cells. GAPDH served as an internal control. D, The western blot analysis of OSR1, SOX9, GSK3b and active b-catenin in A549-siOSR1 cells, A549-siSOX9 cells, A549- siOSR1 + siSOX9 cells and siNC cells. GAPDH served as an internal control. E,F, Representative images of the Matrigel invasion assay and colony formation assay for the H1299-NC, H1299-OSR1, H1299-SOX9 and H1299-OSR1 + SOX9 cells. The cell number for each group is shown in the histogram. G,H, Representative images of the Matrigel invasion assay and colony formation assay for the A549-siNC, A549- siOSR1, A549-siSOX9 and A549-siOSR1 + siSOX9 cells. The cell number for each group is shown in the histogram. *P < .05; **P < .01

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4°C with antibodies against OSR1 (1:150, Santa Cruz Biotechnology), SOX9 (1:150, Santa Cruz Biotechnology), b-catenin (1:100, Santa Cruz Biotechnology), active b-catenin (1:500, Cell Signaling Technology), GSK3b (1:500, Cell Signaling Technology), p-GSK3b (1:500, Cell Signaling Technology), cyclin D1 (1:100, Santa Cruz Biotechnology), c-Myc (1:200, BD Biosciences), MMP7 (1:100, Santa Cruz Biotechnology), LDL receptor related protein 6 (LRP6; 1:100, Santa Cruz Biotechnology), axin (1:100, Santa Cruz Biotechnology), dishevelled segment polarity protein 1 (DVL1; 1:100, Santa Cruz Biotechnology), lymphoid enhancer binding factor 1 (LEF1; 1:100, Santa Cruz Biotechnology), snail (1:500, Cell Signaling Technology), E-cadherin (1:500, Cell Signaling Technology), N-cadherin (1:500, Cell Signaling Technology) and glyceraldeyhde-3-phosphate dehydrogenase (GAPDH; 1:2000, Santa Cruz Biotechnology).

Techniques: Transfection, Western Blot, Control, Invasion Assay, Colony Assay