Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Prolonged Absence of Mechanoluminal Stimulation in Human Intestine Alters the Transcriptome and Intestinal Stem Cell Niche

doi: 10.1016/j.jcmgh.2016.12.008

Figure Lengend Snippet: Primary and Secondary Antibodies

Article Snippet: LGR5 , Rabbit , Miltenyi (San Diego, CA) , 130-104-945 , 1:200.

Techniques: Transduction, Plasmid Preparation

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Prolonged Absence of Mechanoluminal Stimulation in Human Intestine Alters the Transcriptome and Intestinal Stem Cell Niche

doi: 10.1016/j.jcmgh.2016.12.008

Figure Lengend Snippet: qPCR Primer List

Article Snippet: LGR5 , Rabbit , Miltenyi (San Diego, CA) , 130-104-945 , 1:200.

Techniques: Sequencing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Prolonged Absence of Mechanoluminal Stimulation in Human Intestine Alters the Transcriptome and Intestinal Stem Cell Niche

doi: 10.1016/j.jcmgh.2016.12.008

Figure Lengend Snippet: Absence of mechanoluminal stimulation increases LGR5 mRNA expression and downstream Wnt signaling gene expression. ( A and B ) Quantification and in situ hybridization of LGR5 ISCs within the crypts of fed and unfed intestine. Inset : High-magnification photographs label LGR5-positive in situ hybridization with purple arrows . qPCR comparison of ( C ) LGR5 , ( D ) CCND1 , and ( E ) MYC mRNA expression between 6 and 7 pairs of matched fed vs unfed intestine with outliers excluded. ( F ) Representative images of Western blot analysis of STAT3 phosphorylation from 3 pairs of matched tissue. Images were obtained on an upright Leica DM5500B immunofluorescence microscope using Leica Suite Advanced Fluorescence (LAS AF) 6000 software, processed with ImageJ software. Scale bars : 50 μm. * P < .05. Grey bars on plots indicate median with 95% confidence interval.

Article Snippet: LGR5 , Rabbit , Miltenyi (San Diego, CA) , 130-104-945 , 1:200.

Techniques: Expressing, Gene Expression, In Situ Hybridization, Comparison, Western Blot, Phospho-proteomics, Immunofluorescence, Microscopy, Fluorescence, Software

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Prolonged Absence of Mechanoluminal Stimulation in Human Intestine Alters the Transcriptome and Intestinal Stem Cell Niche

doi: 10.1016/j.jcmgh.2016.12.008

Figure Lengend Snippet: Global decrease of LGR5+, SOX9+, and OLFM4+ intestinal stem cell populations occurs in the absence of mechanoluminal flow. ( A–H ) Quantification and immunofluorescence staining of LGR5+ rapidly cycling intestinal stem cells per crypt with proliferating cell nuclear antigen counterstain in fed vs unfed intestine. ( C and F ) LYZ+ Paneth cells with E-cadherin (Ecad) counterstain of same crypt shown in panels A and D , respectively, for comparison. ( I–N ) Quantification and immunofluorescence staining of SOX9 and OLFM4 intestinal stem cells per crypt in fed vs unfed intestine. Images were obtained on an upright Leica DM5500B immunofluorescence microscope using Leica Suite Advanced Fluorescence (LAS AF) 6000 software, processed with ImageJ software. Scale bars : ( B , C , E , and F ) 50 μm, and ( A , D , I , J , L , and M ) 100 μm. * P < .05. Grey bars on plots indicate median with 95% confidence interval. PCNA, proliferating cell nuclear antigen.

Article Snippet: LGR5 , Rabbit , Miltenyi (San Diego, CA) , 130-104-945 , 1:200.

Techniques: Immunofluorescence, Staining, Comparison, Microscopy, Fluorescence, Software

Journal: Translational Cancer Research

Article Title: The biological functions of LGR5 in promoting non-small cell lung cancer progression

doi: 10.21037/tcr.2019.01.24

Figure Lengend Snippet: LGR5 expression in NSCLC tissues. (A) LGR5 expression in NSCLC tissues and matched nontumor lung tissues (P=0.0071); (B) LGR5 expression in NSCLC tumors of different sizes (P=0.2934); (C) LGR5 expression in different lymph node statuses of NSCLC (P=0.1210); (D) LGR5 expression in different TNM stages of NSCLC (P=0.2601). Statistical analysis was performed by a nonparametric t -test. **, P<0.01. LGR5, leucine-rich repeat-containing G-protein-coupled receptor 5; NSCLC, non-small cell lung cancer.

Article Snippet: The expression of LGR5 in transfected A549/H1299 cells was measured using the rat anti-human LGR5 antibody (Miltenyi Biotec, Germany) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Translational Cancer Research

Article Title: The biological functions of LGR5 in promoting non-small cell lung cancer progression

doi: 10.21037/tcr.2019.01.24

Figure Lengend Snippet: Overexpression of LGR5 in human NSCLC cell lines. (A) LGR5 mRNA expression was estimated by qRT-PCR. The expression of LGR5 in A549/LGR5 and H1299/LGR5 cells was higher than that in matched Mock cells (P<0.0001, P=0.0006, respectively); (B) the expression of LGR5 was detected by flow cytometry. Gray = isotype control, black line = expression of LGR5; (C) the stemness-related gene expression was detected in the transfected cells. The expression of NOTCH1 was higher in both A549/LGR5 and H1299/LGR5 cells compared to that in A549/Mock and H1299/Mock cells (P=0.0036, P=0.0192); (D) the expression of NOTCH1 was detected by Western blot in the transfected cells. NOTCH1 expression was increased in LGR5-overexpressing NSCLC cell lines. Statistical analysis by unpaired t -test with Welch correction. ***, P<0.001, ****, P<0.0001. LGR5, Leucine-rich repeat-containing G-protein-coupled receptor 5; NSCLC, non-small cell lung cancer.

Article Snippet: The expression of LGR5 in transfected A549/H1299 cells was measured using the rat anti-human LGR5 antibody (Miltenyi Biotec, Germany) according to the manufacturer’s instructions.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Flow Cytometry, Control, Gene Expression, Transfection, Western Blot

Journal: Translational Cancer Research

Article Title: The biological functions of LGR5 in promoting non-small cell lung cancer progression

doi: 10.21037/tcr.2019.01.24

Figure Lengend Snippet: Overexpression of LGR5 promoted the proliferation of NSCLC cells. (A) Overexpression of LGR5 promoted the proliferation of A549 cells (P=0.0002); (B) overexpression of LGR5 promoted the proliferation of H1299 cells (P=0.004); (C) comparison of the clone-forming ability of each group. LGR5 promoted the clone formation of A549 and H1299 cells (P=0.0059 and P=0.0018, respectively). Statistical analysis by one-way ANOVA with replicate measures or unpaired t -test. **, P<0.01. LGR5, leucine-rich repeat-containing G-protein-coupled receptor 5; NSCLC, non-small cell lung cancer.

Article Snippet: The expression of LGR5 in transfected A549/H1299 cells was measured using the rat anti-human LGR5 antibody (Miltenyi Biotec, Germany) according to the manufacturer’s instructions.

Techniques: Over Expression, Comparison

Journal: Translational Cancer Research

Article Title: The biological functions of LGR5 in promoting non-small cell lung cancer progression

doi: 10.21037/tcr.2019.01.24

Figure Lengend Snippet: LGR5 enhanced the migration of NSCLC cells. A wound-healing assay was used to examine the effect of LGR5 on the migration of A549 (A) and H1299 cells (B). Statistical analysis by unpaired t -test. *, P<0.05; **, P<0.01. LGR5, leucine-rich repeat-containing G-protein-coupled receptor 5; NSCLC, non-small cell lung cancer.

Article Snippet: The expression of LGR5 in transfected A549/H1299 cells was measured using the rat anti-human LGR5 antibody (Miltenyi Biotec, Germany) according to the manufacturer’s instructions.

Techniques: Migration, Wound Healing Assay

Journal: Translational Cancer Research

Article Title: The biological functions of LGR5 in promoting non-small cell lung cancer progression

doi: 10.21037/tcr.2019.01.24

Figure Lengend Snippet: LGR5 increased the invasion ability of NSCLC cells. A Transwell invasion assay was used to examine the effect of LGR5 on the invasion of A549 (A) and H1299 cells (B). Statistical analysis by unpaired t -test. **, P<0.01; ***, P<0.001. LGR5, Leucine-rich repeat-containing G-protein-coupled receptor 5; NSCLC, non-small cell lung cancer.

Article Snippet: The expression of LGR5 in transfected A549/H1299 cells was measured using the rat anti-human LGR5 antibody (Miltenyi Biotec, Germany) according to the manufacturer’s instructions.

Techniques: Transwell Invasion Assay