Journal: Cell metabolism

Article Title: JAK/STAT3-Regulated Fatty Acid β-Oxidation Is Critical for Breast Cancer Stem Cell Self-Renewal and Chemoresistance

doi: 10.1016/j.cmet.2017.11.001

Figure Lengend Snippet: Key Resources Table

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-CD44 Cell Signaling Technology Cat# 3570S; clone 156-3C11 Mouse monoclonal anti-LEPR Sigma Aldrich Cat# SAB1402837-100UG; clone 2H5 Rabbit polyclonal anti-CPT1B Abcam Cat# ab104662 Rabbit polyclonal anti-STAT3 Santa Cruz Biotech Cat# SC-482 Rabbit polyclonal anti-pY705-STAT3 Cell Signaling Technology Cat#9145S Rabbit polyclonal anti-GAPDH Santa Cruz Biotech Cat#sc-25778 Rabbit polyclonal anti-phospho-Jak2 (Tyr1007/1008) Cell Signaling Technology Cat# 3771S RNA polymerase Sigma Aldrich Cat#05-952 Normal rabbit IgG Santa Cruz Biotech Cat# SC-2027 Leptin neutralizing antibody: Mouse monoclonal anti-LEPR R&D Systems, Inc. Cat# AF497 Bacterial and Virus Strains N/A Biological Samples Breast cancer patient specimens City of Hope National Cancer Center N/A Chemicals, Peptides, and Recombinant Proteins Recombinant LEPR BD Pharmingen Cat# 564376 Recombinant CCR2 BD Pharmingen Cat# 561744 Recombinant CXCR1 BD Pharmingen Cat# 555939 Recombinant CXCR2 BD Pharmingen Cat# 551126 [ 3 H]-palmitic acid PerkinElmer Inc Cat# NET043001MC [ 3 H]-Acetyl Coenzyme A PerkinElmer Inc Cat# NET290050UC Acetyl-coA Sigma Aldrich Cat# A2056 Myristic acid Sigma Aldrich Cat# M3128 paclitaxel Sigma Aldrich Cat# T7402 Etomoxir Sigma Aldrich Cat# 236020 Accutase Millipore Cat# SCR005 Critical Commercial Assays Imprint Chromatin-Immunoprecipitation kit Sigma Aldrich Cat# CHP1-96RXN Adipokine array R&D Systems Cat# ARY024 ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set Sigma Aldrich Cat#17-620 RNeasy Plus Mini Kit Qiagen Cat#74134 iScript cDNA Synthesis Kit Bio-Rad Cat#1708890 iQ SYBR Green Supermix Bio-Rad Cat#1708880 Deposited Data N/A Experimental Models: Cell Lines MCF10A ATCC CRL-10317 MCF-7 ATCC HTB-22 MDA-MB-468 ATCC HTB-132 MDA-MB-231-Parental This paper N/A MDA-MB-231-Resistant This paper N/A BBM2 Neman and Jandial, 2004 N/A BBM3 Neman and Jandial, 2004 N/A Hs578T ATCC HTB-126 MDA-MB-436 ATCC HTB-130 HCC1500 ATCC CRL-2329 BT20 ATCC HTB-19 Preadipocyte Zen-Bio BR-F Experimental Models: Organisms/Strains Mouse: FVB/N-Tg(MMTV-PyVT)634Mul/J The Jackson Laboratory Cat# 002374 Oligonucleotides Stat3 siRNA (h) Santa Cruz Biotech Cat# sc-29493 Please see supplemental table 1 for primer sequence Recombinant DNA pRC-CMV-STAT3C This paper N/A pGL 3.1 Promega Cat# E1751 pGL3.1-CPT1B This paper N/A Software and Algorithms Oncomine Rhodes and Chinnaiyan, 2004. https://www.oncomine.org/ Open in a separate window Key Resources Table Inhibition of FAO preferentially eliminates BCSCs JAK/STAT3 activates FAO through transcription of CPT1B Adipocyte-derived leptin is critical for JAK/STAT3-FAO in BCSCs Targeting FAO/leptin inhibits BCSCs, chemoresistance and breast tumor growth

Techniques: Recombinant, Chromatin Immunoprecipitation, SYBR Green Assay, Sequencing, Software

Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: Expression of lncRNA AL355711 and ABCG1 in advanced atherosclerotic plaques compared with normal arterial intimae, as detected using ArrayStar.

Article Snippet: Formalin-fixed, paraffin-embedded sections (4- μ m-thick) of mouse aortic roots were incubated with ABCG1 (overnight at 4°C, 1:1,00; NB400-132SS, Novus Biologicals, LLC) and MMP3 (overnight at 4°C, 1:1,00; sc-21732, Santa Cruz Biotechnology, Inc.) antibodies, and the secondary antibodies using the GTVisionTM Anti-mouse/Anti-rabbit Immunohistochemical Analysis kit (including DAB; HRP conjugates, no dilution, cat. no. GK500710; Gene Tech, Inc.) according to the manufacturer's instructions.

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: lncRNA AL355711 expression is higher in human atherosclerotic plaques. (A) Positional association between lncRNA AL355711 and ABCG1. (B) Expression of lncRNA AL355711 in plaque tissues was significantly increased. Data shown in graphs are the mean ± SD. * P<0.05. (C) Expression of lncRNA AL355711 was significantly increased in circulating leukocytes, as detected by bioinformatics analysis in GSE12288 [RNA was extracted from 110 patients with CAD (CADi >23) and from 112 partially matched controls without CAD (CADi=0)]. Gene expression was assessed using Affymetrix U133A chips (P=0.0002497; <0.001). (D) Expression of ABCG1 was significantly increased in circulating leukocytes, as detected using bioinformatics analysis of the GSE12288 dataset (P=0.0005683; <0.001). (E) Correlation between AL355711 and ABCG1 RNA levels in circulating leukocytes was determined using Pearson's correlation analysis using the GSE12288 dataset (Pearson's R=0.15; P=0.027; <0.05). (F) lncRNA AL355711 was successfully knocked down, and the mRNA level of ABCG1 decreased significantly. Data shown in the graphs are the mean ± SD. * P<0.05. (G) Protein level of ABCG1 was significantly decreased following lncRNA AL355711 knockdown. Data shown in the graphs are the mean ± SD. * P<0.05. lncRNA, long non-coding RNA; ABCG1, ATP-binding cassette sub-family G member 1; CAD, coronary artery disease.

Article Snippet: Formalin-fixed, paraffin-embedded sections (4- μ m-thick) of mouse aortic roots were incubated with ABCG1 (overnight at 4°C, 1:1,00; NB400-132SS, Novus Biologicals, LLC) and MMP3 (overnight at 4°C, 1:1,00; sc-21732, Santa Cruz Biotechnology, Inc.) antibodies, and the secondary antibodies using the GTVisionTM Anti-mouse/Anti-rabbit Immunohistochemical Analysis kit (including DAB; HRP conjugates, no dilution, cat. no. GK500710; Gene Tech, Inc.) according to the manufacturer's instructions.

Techniques: Expressing, Gene Expression, Knockdown, Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: lncRNA AL355711 regulates MMP3 expression through the ABCG1 pathway. (A) Expression of MMP3 was significantly increased, as detected using bioinformatics analysis of the GSE12288 dataset. P=0.0000193; <0.001. (B) Correlation between AL355711 and MMP3 RNA levels in circulating leukocytes identifies patients with coronary artery disease using Pearson's correlation analysis of the GSE12288 dataset (Pearson's R=0.21; P=0.00002; <0.05). (C) Correlation between ABCG1 and MMP3 mRNA levels in circulating leukocytes, as detected using Pearson's correlation analysis of the GSE12288 dataset (Pearson's R=0.32; P=0.000000012; <0.05). (D) Correlation heatmap of AL355711, ABCG1 and MMP3 RNA levels in circulating leukocytes was drawn using Pearson's correlation analysis of the GSE12288 dataset. (E) Transfection with si-ABCG1 was successful, and the scrambled siRNA negative control had no effect on ABCG1 expression. Data shown in the graphs are the mean ± SD. * P<0.05; ns, indicates no significant differences. (F) Knockdown of ABCG1 enhanced the downregulation of ABCG1 by lncRNA AL355711. Data shown in the graphs are the mean ± SD. * P<0.05; ns, indicates no significant differences. lncRNA, long non-coding RNA; ABCG1, ATP-binding cassette sub-family G member 1; MMP3, matrix metalloproteinase 3.

Article Snippet: Formalin-fixed, paraffin-embedded sections (4- μ m-thick) of mouse aortic roots were incubated with ABCG1 (overnight at 4°C, 1:1,00; NB400-132SS, Novus Biologicals, LLC) and MMP3 (overnight at 4°C, 1:1,00; sc-21732, Santa Cruz Biotechnology, Inc.) antibodies, and the secondary antibodies using the GTVisionTM Anti-mouse/Anti-rabbit Immunohistochemical Analysis kit (including DAB; HRP conjugates, no dilution, cat. no. GK500710; Gene Tech, Inc.) according to the manufacturer's instructions.

Techniques: Expressing, Transfection, Negative Control, Knockdown, Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: ABCG1 and MMP3 are highly expressed in an animal model of atherosclerosis. An animal model of atherosclerosis was successfully constructed by feeding 6-week-old apolipoprotein E knockout mice with a Western HFD for 12 weeks. (A) Representative en face images of the ascending aorta stained with Oil Red O (×2.5 magnification). A blue mouse pad was used as the supporting platform for the ascending aortas (hence the green-like background). Data shown in the graphs are mean ± SD. * P<0.05. In total, 4 mice were used as the controls and 5 mice were used in the HFD group. The experiments were performed once. (B) Aortic root cross-sections stained with H&E. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (C) Aortic root cross-sections stained with Masson's stain indicated that the HFD diet significantly reduced the collagen fiber content. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (D) ABCG1 immunohistochemical staining revealed that the HFD significantly increased the ABCG1 levels in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (E) MMP3 immunohistochemical staining revealed that the HFD significantly increased the level of ABCG1 in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. Scale bar left panel, 250 µ m (×100 magnification); scale bar right panel, 100 µ m (×200 magnification). The images on the right panel are an enlarge magnification of the red boxed areas of the images in the left panels. (F) MMP3 immunohistochemical staining revealed that the HFD significantly increased the level of ABCG1 in plaques. Data shown in the graphs are the mean ± SD. * P<0.05. The same paraffin-embedded tissue sections were stained for MMP3 and ABCG1, both of which were highly expressed in the same section of atherosclerotic plaques. Scale bar left panel, 100 µ m (×200 magnification); arrows indicate the red boxed areas, which are presented as an enlarged magnification in the images on the right (×20,000 magnification). lncRNA, long non-coding RNA; ABCG1, ATP-binding cassette sub-family G member 1; MMP3, matrix metalloproteinase 3; HFD, high-fat diet; H&E, hematoxylin and eosin.

Article Snippet: Formalin-fixed, paraffin-embedded sections (4- μ m-thick) of mouse aortic roots were incubated with ABCG1 (overnight at 4°C, 1:1,00; NB400-132SS, Novus Biologicals, LLC) and MMP3 (overnight at 4°C, 1:1,00; sc-21732, Santa Cruz Biotechnology, Inc.) antibodies, and the secondary antibodies using the GTVisionTM Anti-mouse/Anti-rabbit Immunohistochemical Analysis kit (including DAB; HRP conjugates, no dilution, cat. no. GK500710; Gene Tech, Inc.) according to the manufacturer's instructions.

Techniques: Animal Model, Construct, Knock-Out, Western Blot, Staining, Immunohistochemical staining, Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: Long non-coding RNA AL355711 promotes smooth muscle cell migration through the ABCG1/MMP3 pathway

doi: 10.3892/ijmm.2021.5040

Figure Lengend Snippet: Schematic diagram depicting the role of lncRNA AL355711 in the regulation of smooth muscle cell migration. lncRNA AL355711 promoted the transcription of ABCG1, which promoted the protein expression of MMP3, degraded the extracellular matrix, and promoted smooth muscle cell migration. lncRNA, long non-coding RNA; ABCG1, ATP-binding cassette sub-family G member 1; MMP3, matrix metalloproteinase 3.

Article Snippet: Formalin-fixed, paraffin-embedded sections (4- μ m-thick) of mouse aortic roots were incubated with ABCG1 (overnight at 4°C, 1:1,00; NB400-132SS, Novus Biologicals, LLC) and MMP3 (overnight at 4°C, 1:1,00; sc-21732, Santa Cruz Biotechnology, Inc.) antibodies, and the secondary antibodies using the GTVisionTM Anti-mouse/Anti-rabbit Immunohistochemical Analysis kit (including DAB; HRP conjugates, no dilution, cat. no. GK500710; Gene Tech, Inc.) according to the manufacturer's instructions.

Techniques: Migration, Expressing, Binding Assay