mtbp  (Proteintech)

Journal: Cancers

Article Title: Characterization of an Mtbp Hypomorphic Allele in a Diethylnitrosamine-Induced Liver Carcinogenesis Model

doi: 10.3390/cancers15184596

Figure Lengend Snippet: Generation and characterization of Mtbp H/− mice. ( a ) Genomic organization of the murine Mtbp gene, Mtbp H targeting vector, and targeted allele. The vector is constructed as a conditional allele using two different recombinase systems, Flp/Frt and Cre/loxP. The hyg cassette ( TKhygpA ) attenuates transcription of the endogenous Mtbp gene, resulting in the Mtbp H allele. Representative results of Southern blotting ( bottom , left ) following Eco RV restriction enzyme digestion of the genomic DNA from ES cell clones (#102, 103, 114) using the 5′ and 3′ probe set in exon 2 and exon 9, respectively. Genomic PCR ( bottom , right ) using the genomic DNA from mice (#7, 8) with primers of I6F and E7R, showing successful germline transmission. ( b ) Results of qRT-PCR for Mtbp using mRNA from Mtbp +/+ and Mtbp H/− mouse livers. Data are normalized with values of Gapdh mRNA. Error bars: means + S.E. from three independent experiments. Student’s t test: **, p < 0.01. ( c ) Western blotting for Mtbp and Gapdh using protein extracts from liver tissues isolated from Mtbp +/+ and Mtbp H/− mice. ( d ) IHC for Mtbp using liver tissues from Mtbp +/+ and Mtbp H/− mice (2 representative images from each genotype). Scale bar, 25 μm. The uncropped blots are shown in .

Article Snippet: The mouse mRNA expression for Mtbp and Gapdh was analyzed with quantitative RT-PCR (qRT-PCR) with TaqMan probes for Mtbp (Mm00519571_m1_g1 , Thermo Fisher Scientific, Waltham, MA, USA) and Gapdh (Mm99999915_g1 ) using Applied Biosystems ViiA7 (Life Technologies, Carlsbad, CA, USA).

Techniques: Plasmid Preparation, Construct, Southern Blot, Clone Assay, Transmission Assay, Quantitative RT-PCR, Western Blot, Isolation

Journal: Cancers

Article Title: Characterization of an Mtbp Hypomorphic Allele in a Diethylnitrosamine-Induced Liver Carcinogenesis Model

doi: 10.3390/cancers15184596

Figure Lengend Snippet: Increased cell migration of Mtbp H/− MEFs. ( a ) Results of qRT-PCR for Mtbp using mRNA from Mtbp +/+ , Mtbp +/− , and Mtbp H/− MEFs. Data are normalized with values of Gapdh mRNA. Error bars: means ± S.E. from three independent experiments. Student’s t test: **, p < 0.01. ( b ) 3T3 assays using Mtbp +/+ and Mtbp H/− MEFs. Error bars: means + S.E. from three independent experiments. Student’s t test: not significant. ( c ) Transwell migration assays using Mtbp +/+ , Mtbp +/− , and Mtbp H/− MEFs. Cells were plated on the upper chambers of the Transwell. Migrating cells in the entire fields were counted 14 h later. A summary graph ( top ) and representative images ( bottom ). Scale bar, 25 μm. Error bars: means ± S.E. from three independent experiments. Student’s t test: *, p < 0.05. ( d ) Immunofluorescence studies for p-Erk following treatment with vehicle (EGF-) or 50 ng/mL of EGF (EGF+) for 30 min using Mtbp +/+ and Mtbp H/− MEFs. Scale bar, 25 μm. Graph showing the number of cells with different Mtbp locations (Nucleus ≥ Cytoplasm or Nucleus < Cytoplasm) in the absence of EGF treatment ( n = 50). Fisher’s exact test (two tailed): **, p < 0.01.

Article Snippet: The mouse mRNA expression for Mtbp and Gapdh was analyzed with quantitative RT-PCR (qRT-PCR) with TaqMan probes for Mtbp (Mm00519571_m1_g1 , Thermo Fisher Scientific, Waltham, MA, USA) and Gapdh (Mm99999915_g1 ) using Applied Biosystems ViiA7 (Life Technologies, Carlsbad, CA, USA).

Techniques: Migration, Quantitative RT-PCR, Immunofluorescence, Two Tailed Test

Journal: Cancers

Article Title: Characterization of an Mtbp Hypomorphic Allele in a Diethylnitrosamine-Induced Liver Carcinogenesis Model

doi: 10.3390/cancers15184596

Figure Lengend Snippet: Increased liver carcinogenesis in DEN-treated Mtbp H/− mice. ( a ) Schematic of DEN-induced liver carcinogenesis studies. Mice were injected with DEN at 25 mg/kg of body weight two weeks after birth. ( b ) Representative images of the liver ( top ) and lung ( bottom ) of a non-DEN-treated Mtbp +/+ mouse and DEN-treated Mtbp +/+ , Mtbp +/− , and Mtbp H/− mice with tumors. White and Black arrows indicate tumor nodules in liver and lung, respectively. ( c ) Kaplan–Meier survival curves of Mtbp +/+ ( n = 21) , Mtbp +/− ( n = 18) , and Mtbp H/− ( n = 18) mice injected with DEN. Log-rank test: * p < 0.05; ns: not significant. ( d ) Numbers of mice with liver nodules in Mtbp +/+ ( n = 21) and Mtbp H/− ( n = 18) mice. Fisher’s exact test (two tailed); ns: not significant. ( e ) IHC for p-Erk using HCC tumors from Mtbp +/+ and Mtbp H/− mice as well as a lung adenoma from a Mtbp +/+ mouse or a lung adenocarcinoma from a Mtbp H/− mouse. Scale bar, 25 μm.

Article Snippet: The mouse mRNA expression for Mtbp and Gapdh was analyzed with quantitative RT-PCR (qRT-PCR) with TaqMan probes for Mtbp (Mm00519571_m1_g1 , Thermo Fisher Scientific, Waltham, MA, USA) and Gapdh (Mm99999915_g1 ) using Applied Biosystems ViiA7 (Life Technologies, Carlsbad, CA, USA).

Techniques: Injection, Two Tailed Test

Journal: Cancers

Article Title: Characterization of an Mtbp Hypomorphic Allele in a Diethylnitrosamine-Induced Liver Carcinogenesis Model

doi: 10.3390/cancers15184596

Figure Lengend Snippet: DEN-induced lung tumors and metastases from the liver in Mtbp +/+ and Mtbp H/ − mice.

Article Snippet: The mouse mRNA expression for Mtbp and Gapdh was analyzed with quantitative RT-PCR (qRT-PCR) with TaqMan probes for Mtbp (Mm00519571_m1_g1 , Thermo Fisher Scientific, Waltham, MA, USA) and Gapdh (Mm99999915_g1 ) using Applied Biosystems ViiA7 (Life Technologies, Carlsbad, CA, USA).

Techniques: Mouse Assay

Journal: Clinical and Translational Medicine

Article Title: C9orf142 transcriptionally activates MTBP to drive progression and resistance to CDK4/6 inhibitor in triple‐negative breast cancer

doi: 10.1002/ctm2.1480

Figure Lengend Snippet: C9orf142 transcriptionally regulates MTBP expression. (A) LM2‐4175 cells stably expressing shNC and shC9orf142 (#1 and #2) were subjected to label‐free quantitative proteomic analysis. The numbers of differentially expressed proteins between cells expressing shNC and shC9orf142 based on the cut‐off value of 1.5‐fold change are shown. (B) Heatmap of the top 30 up‐regulated and down‐regulated proteins in LM2‐4175 cells stably expressing shNC and shC9orf142 (#1 and #2). (C and D) Gene ontology‐biological process (GO‐BP) (C) and GO‐molecular function (GO‐MF) (D) of differentially expressed proteins between cells expressing shNC and shC9orf142. (E) KEGG pathway analysis of differentially expressed proteins between cells expressing shNC and shC9orf142. (F and G) Hs578T and LM2‐4175 cells stably expressing shNC and shC9orf142 (#1 and #2) were subjected to immunoblotting assays with the indicated antibodies (F) and RT‐qPCR analysis (G). (H and I) SUM159 and MDA‐MB‐231 (MDA‐231) cells stably expressing pCDH and Flag‐C9orf142 were subjected to immunoblotting assays with the indicated antibodies (H) and RT‐qPCR analysis (I). * p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significance.

Article Snippet: The specific short hairpin RNA (shRNA) sequences targeting C9orf142 and MTBP were acquired from Sigma‐Aldrich Advanced Genomics ( www.sigmaaldrich.cn ) (Table ).

Techniques: Expressing, Stable Transfection, Western Blot, Quantitative RT-PCR

Journal: Clinical and Translational Medicine

Article Title: C9orf142 transcriptionally activates MTBP to drive progression and resistance to CDK4/6 inhibitor in triple‐negative breast cancer

doi: 10.1002/ctm2.1480

Figure Lengend Snippet: C9orf142 is recruited to the MTBP promoter and enhances its promoter activity. (A) Line diagram showing the regions of the MTBP promoter used for ChIP‐qPCR assays. (B and C) SUM159 and MDA‐MB‐231 cells stably expressing empty vector pCDH or Flag‐C9orf142 were subjected to ChIP assays and followed by RT‐qPCR assays (primer #3 and #4). The ChIP assays were carried out using an anti‐Flag antibody or IgG, where IgG was used as a negative control. Recruitment of Flag‐C9orf142 to the MTBP promoter was normalized to Input. Representative results of primer #1 and #2 are shown in Figure and . (D) Line diagram showing the regions of the MTBP promoter used for dual‐luciferase reporter assays. (E) HEK293T cells stably expressing pCDH or Flag‐C9orf142 were transfected with a luciferase reporter construct encoding pGL3 or pGL3‐MTBP (#1, #2, #3 and #4, respectively). Relative fluorescence activity was normalized to co‐transfected Renilla luciferase. (F) HEK293T cells stably expressing pCDH or Flag‐C9orf142 were transfected with a luciferase reporter construct encoding pGL3 or pGL3‐MTBP (#3 and #4 in combination). Relative fluorescence activity was normalized to co‐transfected Renilla luciferase. (G) HEK293T cells stably expressing shNC or shC9orf142 (#1 and #2) were transfected with a luciferase reporter construct encoding pGL3 or pGL3‐MTBP (#3 and #4 in combination). Relative fluorescence activity was normalized to co‐transfected Renilla luciferase. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significance.

Article Snippet: The specific short hairpin RNA (shRNA) sequences targeting C9orf142 and MTBP were acquired from Sigma‐Aldrich Advanced Genomics ( www.sigmaaldrich.cn ) (Table ).

Techniques: Activity Assay, Stable Transfection, Expressing, Plasmid Preparation, Quantitative RT-PCR, Negative Control, Luciferase, Transfection, Construct, Fluorescence

Journal: Clinical and Translational Medicine

Article Title: C9orf142 transcriptionally activates MTBP to drive progression and resistance to CDK4/6 inhibitor in triple‐negative breast cancer

doi: 10.1002/ctm2.1480

Figure Lengend Snippet: C9orf142 accelerates TNBC progression via regulating MTBP expression. (A) SUM159 and MDA‐MB‐231 cells stably expressing pCDH or Flag‐C9orf142 were transfected with shNC or shMTBP #3. After 48 h of transfection, cells were subjected to immunoblotting analysis with the indicated antibodies. (B and C) SUM159 and MDA‐MB‐231 cells stably expressing pCDH or Flag‐C9orf142 alone or in combination with shNC or shMTBP #3 were subjected to CCK‐8 (B) and colony formation assays (C). Representative images of survival colonies are shown inFigure . (D and E) SUM159 and MDA‐MB‐231 cells stably expressing pCDH or Flag‐C9orf142 alone or in combination with shNC or shMTBP #3 were subjected to Transwell migration assays (D) and Matrigel‐coated invasion assays (E). Representative images of migrated and invaded cells are shown in Figure and . (F–H) A total of 3 × 10 6 MDA‐MB‐231 cells stably expressing expressing pCDH or Flag‐C9orf142 alone or in combination with shNC or shMTBP #3 were inoculated into mammary fat pads of 6‐week‐old BALB/c female nude mice ( n = 10). After 30 days of injection, mice were sacrificed and xenograft tumours were removed. Image of removed xenograft tumours (F), tumour volume (G), and tumour weight (H) are shown. (I‐K) A total of 1×10 6 LM2‐4175 cells stably expressing shNC or shC9orf142 (#1 and #2) were injected into the tail vein of mammary fat pads of 7‐week‐old BALB/c female nude mice ( n = 10). After 6 weeks of injection, mice were sacrificed and lungs were removed. Representative images of lung metastasis (I), representative images of HE staining of lung tissues (J), and the incidence of lung metastasis (K) are shown. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significance.

Article Snippet: The specific short hairpin RNA (shRNA) sequences targeting C9orf142 and MTBP were acquired from Sigma‐Aldrich Advanced Genomics ( www.sigmaaldrich.cn ) (Table ).

Techniques: Expressing, Stable Transfection, Transfection, Western Blot, CCK-8 Assay, Migration, Injection, Staining

Journal: Clinical and Translational Medicine

Article Title: C9orf142 transcriptionally activates MTBP to drive progression and resistance to CDK4/6 inhibitor in triple‐negative breast cancer

doi: 10.1002/ctm2.1480

Figure Lengend Snippet: Knockdown of C9orf142 enhances the sensitivity of TNBC cells to abemaciclib both in vitro and in vivo. (A) Hs578T and LM2‐4175 stably expressing empty vector shNC or shC9orf142 (#1 and #2) were treated with or without increasing doses of abemaciclib. Cell viability was determined using CCK‐8 assays. The IC50 values are shown. (B and C) Hs578T and LM2‐4175 cells stably expressing empty vector shNC or shC9orf142 (#1 and #2) were treated without or with the indicated concentrations of abemaciclib and subjected to colony formation assays. Representative images of survival colonies and corresponding quantitative results are shown in B and C, respectively. (D–F) A total of 1 × 10 6 LM2‐4175 cells stably expressing shNC or shC9orf142 (#1 and #2) were inoculated into mammary fat pads of 6‐week‐old BALB/c female nude mice ( n = 20). After 16 days of injection, each group is randomly divided into two groups ( n = 10), and the mice were administered with or without abemaciclib (25 mg/kg) by oral gavage once daily for 14 consecutive days. After 30 days of injection, mice were sacrificed and xenograft tumours were removed. Image of removed xenograft tumours (D), tumour volume (E) and tumour weight (F) are shown. (G) The proposed working model. C9orf142 transcriptionally activates MTBP and regulates its downstream MDM2/p53/p21 signaling axis and cell cycle transition from G1‐to‐S phase to drive the progression and resistance to CDK4/6 inhibitor in TNBC. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significance.

Article Snippet: The specific short hairpin RNA (shRNA) sequences targeting C9orf142 and MTBP were acquired from Sigma‐Aldrich Advanced Genomics ( www.sigmaaldrich.cn ) (Table ).

Techniques: In Vitro, In Vivo, Stable Transfection, Expressing, Plasmid Preparation, CCK-8 Assay, Injection

Journal: Bulletin of Mathematical Biology

Article Title: Effect of Movement on the Early Phase of an Epidemic

doi: 10.1007/s11538-022-01077-5

Figure Lengend Snippet: Absolute value of the difference between the MTBP and stochastic phase approximations of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\mathbb {P}}_0$$\end{document} P 0 as a function of the threshold \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\hat{I}}$$\end{document} I ^ and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$m^{AB}$$\end{document} m AB (Color figure online)

Article Snippet: Instead, we approximate the probability of extinction using multitype branching process (MTBP) approximation (see Milliken ( ); Tritch and Allen ( )).

Techniques:

Journal: Bulletin of Mathematical Biology

Article Title: Effect of Movement on the Early Phase of an Epidemic

doi: 10.1007/s11538-022-01077-5

Figure Lengend Snippet: Absolute value of the difference between the MTBP and stochastic phase approximations of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\mathbb {P}}_0$$\end{document} P 0 as a function of the threshold \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\hat{I}}$$\end{document} I ^ and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\mathcal {R}}_0^A$$\end{document} R 0 A . In alphabetical order \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\mathcal {R}}_0^B$$\end{document} R 0 B = 0.8, 1.2, 2.5 (Color figure online)

Article Snippet: Instead, we approximate the probability of extinction using multitype branching process (MTBP) approximation (see Milliken ( ); Tritch and Allen ( )).

Techniques: