murine mesenchymal stem cell line c3h10t1 2  (ATCC)


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    Name:
    C3H 10T1 2 Clone 8
    Description:
    Applications This line is a suitable transfection host Host Mus musculus mouse
    Catalog Number:
    CCL-226
    Price:
    None
    Applications:
    This line is a suitable transfection host.
    Host:
    Mus musculus, mouse
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    Structured Review

    ATCC murine mesenchymal stem cell line c3h10t1 2
    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal <t>C3H10T1/2</t> cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p
    Applications This line is a suitable transfection host Host Mus musculus mouse
    https://www.bioz.com/result/murine mesenchymal stem cell line c3h10t1 2/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine mesenchymal stem cell line c3h10t1 2 - by Bioz Stars, 2021-06
    86/100 stars

    Images

    1) Product Images from "Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition"

    Article Title: Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition

    Journal: bioRxiv

    doi: 10.1101/2021.01.19.427285

    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p
    Figure Legend Snippet: TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p

    Techniques Used: Inhibition, In Vitro, In Vivo, Luciferase, Concentration Assay, SDS Page, Expressing, Autoradiography, Functional Assay, Transfection

    the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p
    Figure Legend Snippet: the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p

    Techniques Used: Mutagenesis, Luciferase, In Vitro, SDS Page, In Vivo, Construct, Transfection, Expressing

    2) Product Images from "Divergent activities of osteogenic BMP2, and tenogenic BMP12 and BMP13 independent of receptor binding affinities"

    Article Title: Divergent activities of osteogenic BMP2, and tenogenic BMP12 and BMP13 independent of receptor binding affinities

    Journal: Growth Factors (Chur, Switzerland)

    doi: 10.3109/08977194.2011.593178

    BMP activity in C3H10T1/2 cells. (a) C3H10T1/2 cells infected with BRE-Luc construct were treated with increasing concentration of rhBMP2, rhBMP12, or rhBMP13. (b) ALP activity in C3H10T1/2 cells following 2-day incubation with 0, 1, 10, or 100nM of rhBMP2, rhBMP12, or rhBMP13.
    Figure Legend Snippet: BMP activity in C3H10T1/2 cells. (a) C3H10T1/2 cells infected with BRE-Luc construct were treated with increasing concentration of rhBMP2, rhBMP12, or rhBMP13. (b) ALP activity in C3H10T1/2 cells following 2-day incubation with 0, 1, 10, or 100nM of rhBMP2, rhBMP12, or rhBMP13.

    Techniques Used: Activity Assay, Infection, Construct, Concentration Assay, ALP Assay, Incubation

    Induction of SMAD signaling by BMPs in C3H10T1/2 cells. Cell lysates from rhBMP2, rhBMP12, or rhBMP13 treated C3H10T1/2 cells were analyzed by western blot with antibodies to phosphorylated SMAD 1/5/8 and to β-actin or tubulin. Cells were treated with (a)10 nM BMPs for 0 to 30 min, (b) 10 nM BMPs for 0 to 120 min or (c) treated with 100 nM BMPs for 0 to 3 h.
    Figure Legend Snippet: Induction of SMAD signaling by BMPs in C3H10T1/2 cells. Cell lysates from rhBMP2, rhBMP12, or rhBMP13 treated C3H10T1/2 cells were analyzed by western blot with antibodies to phosphorylated SMAD 1/5/8 and to β-actin or tubulin. Cells were treated with (a)10 nM BMPs for 0 to 30 min, (b) 10 nM BMPs for 0 to 120 min or (c) treated with 100 nM BMPs for 0 to 3 h.

    Techniques Used: Western Blot

    Expression of bone, tendon, and cartilage markers by BMPs in C3H10T1/2 cells. mRNA levels were determined by Taqman for the bone markers (a) Ocn (osteocalcin), (b) Runx2 (runt-related transcription factor 2, (c) Alpl (ALP), and (d) Sp7 (osterix), the tendon markers (e) Thbs4 (thrombospondin 4) and (f) Tnmd , (tenomodulin) and the cartilage markers (g) Sox9 (SRY-box containing gene 9), (h) Col2a1 (collagen, type II, alpha 1), (i) Acan (Aggrecan 1) and (j) Col11a1 (collagen, type 11, alpha 1).
    Figure Legend Snippet: Expression of bone, tendon, and cartilage markers by BMPs in C3H10T1/2 cells. mRNA levels were determined by Taqman for the bone markers (a) Ocn (osteocalcin), (b) Runx2 (runt-related transcription factor 2, (c) Alpl (ALP), and (d) Sp7 (osterix), the tendon markers (e) Thbs4 (thrombospondin 4) and (f) Tnmd , (tenomodulin) and the cartilage markers (g) Sox9 (SRY-box containing gene 9), (h) Col2a1 (collagen, type II, alpha 1), (i) Acan (Aggrecan 1) and (j) Col11a1 (collagen, type 11, alpha 1).

    Techniques Used: Expressing, ALP Assay

    3) Product Images from "The Osteogenic and Tenogenic Differentiation Potential of C3H10T1/2 (Mesenchymal Stem Cell Model) Cultured on PCL/PLA Electrospun Scaffolds in the Absence of Specific Differentiation Medium"

    Article Title: The Osteogenic and Tenogenic Differentiation Potential of C3H10T1/2 (Mesenchymal Stem Cell Model) Cultured on PCL/PLA Electrospun Scaffolds in the Absence of Specific Differentiation Medium

    Journal: Materials

    doi: 10.3390/ma10121387

    Fluorescence microscopy observations of C3H10T1/2 cells cultured for 96 h on PCL1000nm ( A , B ) and PCLaligned ( C , D ) scaffolds. ( A , C ) Actin filaments visualized with rhodamine-phalloidine ( red ) and nuclei visualized with DAPI ( blue ). ( B , D ) higher magnifications of nuclei (DAPI) of region of interest (white rectangles in A , C ).
    Figure Legend Snippet: Fluorescence microscopy observations of C3H10T1/2 cells cultured for 96 h on PCL1000nm ( A , B ) and PCLaligned ( C , D ) scaffolds. ( A , C ) Actin filaments visualized with rhodamine-phalloidine ( red ) and nuclei visualized with DAPI ( blue ). ( B , D ) higher magnifications of nuclei (DAPI) of region of interest (white rectangles in A , C ).

    Techniques Used: Fluorescence, Microscopy, Cell Culture

    Fluorescence microscopy ( A , B , E , F , I , J ) and SEM observations ( C , D , G , H , K , L ) of C3H10T1/2 cells cultured for 96 h on PCL60nm ( A , C ), PCL1000nm ( B , D ), PLA ( E , G ), PCLaligned ( F , H ), BlendPLAout ( I , K ) and BlendPCLout ( J , L ) electrospun scaffolds. Fluorescence: Living cells (Calcein AM, green) and dead cell nuclei (EthD-1, red), scale bars 250 µm. SEM: scale bars 100 µm. Solid arrows: continuous cell tissue. Dashed arrows: fibers network visible due to cracks in the cell tissue or to a lower cell density.
    Figure Legend Snippet: Fluorescence microscopy ( A , B , E , F , I , J ) and SEM observations ( C , D , G , H , K , L ) of C3H10T1/2 cells cultured for 96 h on PCL60nm ( A , C ), PCL1000nm ( B , D ), PLA ( E , G ), PCLaligned ( F , H ), BlendPLAout ( I , K ) and BlendPCLout ( J , L ) electrospun scaffolds. Fluorescence: Living cells (Calcein AM, green) and dead cell nuclei (EthD-1, red), scale bars 250 µm. SEM: scale bars 100 µm. Solid arrows: continuous cell tissue. Dashed arrows: fibers network visible due to cracks in the cell tissue or to a lower cell density.

    Techniques Used: Fluorescence, Microscopy, Cell Culture, Proximity Ligation Assay, Ethidium Homodimer Assay

    Related Articles

    Expressing:

    Article Title: Identification of zinc finger protein Bcl6 as a novel regulator of early adipose commitment
    Article Snippet: Furthermore, to establish the direct transcriptional effects of Bcl6 on these overlapping genes, we identified six key factors which have potential Bcl6 binding sites on their promoter through bioinformatics analysis (electronic supplementary material, table S5). .. Among these six targets, STAT1 was of particular interest to us, since RNAi-mediated knockdown of STAT1 expression significantly inhibited adipogenic phenotype more obviously than knockdown of others genes in C3H10T1/2 cells, an effect that mimicked that of PPARγ knockdown ( e ,f ). .. Bcl6 transactivates STAT1 by directly binding to the STAT1 promoter As shown in a ,b , qPCR and western blot analysis in C3H10T1/2 cells confirmed that Bcl6 exerted strong positive effects on STAT1 expression, whereas reducing the Bcl6 expression had the converse effect.

    Cell Culture:

    Article Title: IGF1 potentiates BMP9-induced osteogenic differentiation in mesenchymal stem cells through the enhancement of BMP/Smad signaling
    Article Snippet: Taken together, the findings of this investigation strongly suggest that IGF1 is an excellent candidate enhancer for BMP9-induced osteogenic differentiation in MSCs, which may be mediated by the promotion of BMP9-initialized activation of BMP/Smad signaling transduction. .. Cell culture and chemicals HEK293, C3H10T1/2, and C2C12 cells were purchased from ATCC (VA, USA). .. Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37℃ in 5% CO2 .

    Article Title: SCARA5 plays a critical role in the commitment of mesenchymal stem cells to adipogenesis
    Article Snippet: .. Cell culture and Induction of differentiation A33 and C3H10T1/2 cells (American Type Culture Collection, Bethesda, MD) were propagated and differentiated as described . .. A33 cells were cultured and differentiated into adipocytes as described previously .

    Real-time Polymerase Chain Reaction:

    Article Title: Identification of zinc finger protein Bcl6 as a novel regulator of early adipose commitment
    Article Snippet: STAT1 is required for adipogenesis of C3H10T1/2 cells Since STAT1 was shown to be a target of Bcl6 during adipogenesis, its principal role was further evaluated by its knockdown in C3H10T1/2 cells. .. The efficacy of the RNAi to knockdown STAT1 in C3H10T1/2 cells was confirmed by qPCR and western blot, respectively ( a ,b ). .. At day 8 post-induction, oil red-O staining revealed the dramatic impairment in adipocytic phenotypes for individual siRNAs and almost completely abolished differentiation when siSTAT1-3 was used ( c ).

    Western Blot:

    Article Title: Identification of zinc finger protein Bcl6 as a novel regulator of early adipose commitment
    Article Snippet: STAT1 is required for adipogenesis of C3H10T1/2 cells Since STAT1 was shown to be a target of Bcl6 during adipogenesis, its principal role was further evaluated by its knockdown in C3H10T1/2 cells. .. The efficacy of the RNAi to knockdown STAT1 in C3H10T1/2 cells was confirmed by qPCR and western blot, respectively ( a ,b ). .. At day 8 post-induction, oil red-O staining revealed the dramatic impairment in adipocytic phenotypes for individual siRNAs and almost completely abolished differentiation when siSTAT1-3 was used ( c ).

    Modification:

    Article Title: A Network of Transcription Factors Operates during Early Tooth Morphogenesis
    Article Snippet: The sequence data from each clone were conceptually translated in the reading frame of the library vector, and the sequence identity was determined by searching the mouse genome database at . .. C3H10T1/2 cells (CCL-226; ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; Invitrogen) supplemented with 10% (vol/vol) fetal bovine serum at 37°C in a 5% CO2 humidified atmosphere. .. The cells, in 60-mm dishes, were cotransfected with plasmids encoding Dlx2, Lhx6, Lhx8, Snail, Sp3, or Lef1 and pCMV-FLAG-Msx1 or with plasmids encoding Dlx2, Lhx6, Lhx8, Snail, Sp3, or Lef1 and pCMV-Tag2B (Stratagene) using FuGene 6 reagent (Roche) according to the manufacturer's instructions.

    other:

    Article Title: Identification of zinc finger protein Bcl6 as a novel regulator of early adipose commitment
    Article Snippet: These results indicate that STAT1 is required for the adipogenesis of C3H10T1/2 cells.

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  • 86
    ATCC murine mesenchymal stem cell line c3h10t1 2
    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal <t>C3H10T1/2</t> cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p
    Murine Mesenchymal Stem Cell Line C3h10t1 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine mesenchymal stem cell line c3h10t1 2/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine mesenchymal stem cell line c3h10t1 2 - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    97
    ATCC murine mesenchymal stem cell line c3h10t1 2 the c3h10t1 2
    The effects of Sirt1 pharmacologic activation and inhibition on adipogenic markers in C3HT101/2 cells (A) . Oil-red-o staining in <t>C3H10T1/2</t> cells induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (B,C) Gene expression analysis of adipocyte (B) and mitochondrial markers (C) induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (D) Gene expression analysis of adipocyte markers induced to adipogenesis and supplemented with Ex527 or vehicle (DMSO). Results are Mean ± SEM. * P
    Murine Mesenchymal Stem Cell Line C3h10t1 2 The C3h10t1 2, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/murine mesenchymal stem cell line c3h10t1 2 the c3h10t1 2/product/ATCC
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    murine mesenchymal stem cell line c3h10t1 2 the c3h10t1 2 - by Bioz Stars, 2021-06
    97/100 stars
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    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p

    Journal: bioRxiv

    Article Title: Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition

    doi: 10.1101/2021.01.19.427285

    Figure Lengend Snippet: TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p

    Article Snippet: Cell lines Two cell lines were used for our in vivo assays: Human embryonic kidney cell line HEK293FT (ATCC® ) and murine mesenchymal stem cell line C3H10T1/2 (clone 8, ATCC® CCL26).

    Techniques: Inhibition, In Vitro, In Vivo, Luciferase, Concentration Assay, SDS Page, Expressing, Autoradiography, Functional Assay, Transfection

    the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p

    Journal: bioRxiv

    Article Title: Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition

    doi: 10.1101/2021.01.19.427285

    Figure Lengend Snippet: the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p

    Article Snippet: Cell lines Two cell lines were used for our in vivo assays: Human embryonic kidney cell line HEK293FT (ATCC® ) and murine mesenchymal stem cell line C3H10T1/2 (clone 8, ATCC® CCL26).

    Techniques: Mutagenesis, Luciferase, In Vitro, SDS Page, In Vivo, Construct, Transfection, Expressing

    BMP activity in C3H10T1/2 cells. (a) C3H10T1/2 cells infected with BRE-Luc construct were treated with increasing concentration of rhBMP2, rhBMP12, or rhBMP13. (b) ALP activity in C3H10T1/2 cells following 2-day incubation with 0, 1, 10, or 100nM of rhBMP2, rhBMP12, or rhBMP13.

    Journal: Growth Factors (Chur, Switzerland)

    Article Title: Divergent activities of osteogenic BMP2, and tenogenic BMP12 and BMP13 independent of receptor binding affinities

    doi: 10.3109/08977194.2011.593178

    Figure Lengend Snippet: BMP activity in C3H10T1/2 cells. (a) C3H10T1/2 cells infected with BRE-Luc construct were treated with increasing concentration of rhBMP2, rhBMP12, or rhBMP13. (b) ALP activity in C3H10T1/2 cells following 2-day incubation with 0, 1, 10, or 100nM of rhBMP2, rhBMP12, or rhBMP13.

    Article Snippet: Cell culture conditions The murine mesenchymal stem cell line C3H10T1/2 (clone 8) was purchased from ATCC (CCL-226; Manassas, VA, USA).

    Techniques: Activity Assay, Infection, Construct, Concentration Assay, ALP Assay, Incubation

    Induction of SMAD signaling by BMPs in C3H10T1/2 cells. Cell lysates from rhBMP2, rhBMP12, or rhBMP13 treated C3H10T1/2 cells were analyzed by western blot with antibodies to phosphorylated SMAD 1/5/8 and to β-actin or tubulin. Cells were treated with (a)10 nM BMPs for 0 to 30 min, (b) 10 nM BMPs for 0 to 120 min or (c) treated with 100 nM BMPs for 0 to 3 h.

    Journal: Growth Factors (Chur, Switzerland)

    Article Title: Divergent activities of osteogenic BMP2, and tenogenic BMP12 and BMP13 independent of receptor binding affinities

    doi: 10.3109/08977194.2011.593178

    Figure Lengend Snippet: Induction of SMAD signaling by BMPs in C3H10T1/2 cells. Cell lysates from rhBMP2, rhBMP12, or rhBMP13 treated C3H10T1/2 cells were analyzed by western blot with antibodies to phosphorylated SMAD 1/5/8 and to β-actin or tubulin. Cells were treated with (a)10 nM BMPs for 0 to 30 min, (b) 10 nM BMPs for 0 to 120 min or (c) treated with 100 nM BMPs for 0 to 3 h.

    Article Snippet: Cell culture conditions The murine mesenchymal stem cell line C3H10T1/2 (clone 8) was purchased from ATCC (CCL-226; Manassas, VA, USA).

    Techniques: Western Blot

    Expression of bone, tendon, and cartilage markers by BMPs in C3H10T1/2 cells. mRNA levels were determined by Taqman for the bone markers (a) Ocn (osteocalcin), (b) Runx2 (runt-related transcription factor 2, (c) Alpl (ALP), and (d) Sp7 (osterix), the tendon markers (e) Thbs4 (thrombospondin 4) and (f) Tnmd , (tenomodulin) and the cartilage markers (g) Sox9 (SRY-box containing gene 9), (h) Col2a1 (collagen, type II, alpha 1), (i) Acan (Aggrecan 1) and (j) Col11a1 (collagen, type 11, alpha 1).

    Journal: Growth Factors (Chur, Switzerland)

    Article Title: Divergent activities of osteogenic BMP2, and tenogenic BMP12 and BMP13 independent of receptor binding affinities

    doi: 10.3109/08977194.2011.593178

    Figure Lengend Snippet: Expression of bone, tendon, and cartilage markers by BMPs in C3H10T1/2 cells. mRNA levels were determined by Taqman for the bone markers (a) Ocn (osteocalcin), (b) Runx2 (runt-related transcription factor 2, (c) Alpl (ALP), and (d) Sp7 (osterix), the tendon markers (e) Thbs4 (thrombospondin 4) and (f) Tnmd , (tenomodulin) and the cartilage markers (g) Sox9 (SRY-box containing gene 9), (h) Col2a1 (collagen, type II, alpha 1), (i) Acan (Aggrecan 1) and (j) Col11a1 (collagen, type 11, alpha 1).

    Article Snippet: Cell culture conditions The murine mesenchymal stem cell line C3H10T1/2 (clone 8) was purchased from ATCC (CCL-226; Manassas, VA, USA).

    Techniques: Expressing, ALP Assay

    Fluorescence microscopy observations of C3H10T1/2 cells cultured for 96 h on PCL1000nm ( A , B ) and PCLaligned ( C , D ) scaffolds. ( A , C ) Actin filaments visualized with rhodamine-phalloidine ( red ) and nuclei visualized with DAPI ( blue ). ( B , D ) higher magnifications of nuclei (DAPI) of region of interest (white rectangles in A , C ).

    Journal: Materials

    Article Title: The Osteogenic and Tenogenic Differentiation Potential of C3H10T1/2 (Mesenchymal Stem Cell Model) Cultured on PCL/PLA Electrospun Scaffolds in the Absence of Specific Differentiation Medium

    doi: 10.3390/ma10121387

    Figure Lengend Snippet: Fluorescence microscopy observations of C3H10T1/2 cells cultured for 96 h on PCL1000nm ( A , B ) and PCLaligned ( C , D ) scaffolds. ( A , C ) Actin filaments visualized with rhodamine-phalloidine ( red ) and nuclei visualized with DAPI ( blue ). ( B , D ) higher magnifications of nuclei (DAPI) of region of interest (white rectangles in A , C ).

    Article Snippet: The murine mesenchymal stem cell line C3H10T1/2 [ ] (ATCC CCL-26) initially proliferated in culture flasks at 37 °C, 5% CO2 , in 1 g/L glucose DMEM medium (Sigma-Aldrich, St. Louis, MO, USA) complemented with 10% of Fetal Bovine Serum (Gibco Invitrogen, Waltham, MA USA), 2% of penicillin-streptomycin (Gibco Invitrogen, Waltham, MA, USA) and 2% of 200 mM l -glutamine (Gibco Invitrogen, Waltham, MA, USA).

    Techniques: Fluorescence, Microscopy, Cell Culture

    Fluorescence microscopy ( A , B , E , F , I , J ) and SEM observations ( C , D , G , H , K , L ) of C3H10T1/2 cells cultured for 96 h on PCL60nm ( A , C ), PCL1000nm ( B , D ), PLA ( E , G ), PCLaligned ( F , H ), BlendPLAout ( I , K ) and BlendPCLout ( J , L ) electrospun scaffolds. Fluorescence: Living cells (Calcein AM, green) and dead cell nuclei (EthD-1, red), scale bars 250 µm. SEM: scale bars 100 µm. Solid arrows: continuous cell tissue. Dashed arrows: fibers network visible due to cracks in the cell tissue or to a lower cell density.

    Journal: Materials

    Article Title: The Osteogenic and Tenogenic Differentiation Potential of C3H10T1/2 (Mesenchymal Stem Cell Model) Cultured on PCL/PLA Electrospun Scaffolds in the Absence of Specific Differentiation Medium

    doi: 10.3390/ma10121387

    Figure Lengend Snippet: Fluorescence microscopy ( A , B , E , F , I , J ) and SEM observations ( C , D , G , H , K , L ) of C3H10T1/2 cells cultured for 96 h on PCL60nm ( A , C ), PCL1000nm ( B , D ), PLA ( E , G ), PCLaligned ( F , H ), BlendPLAout ( I , K ) and BlendPCLout ( J , L ) electrospun scaffolds. Fluorescence: Living cells (Calcein AM, green) and dead cell nuclei (EthD-1, red), scale bars 250 µm. SEM: scale bars 100 µm. Solid arrows: continuous cell tissue. Dashed arrows: fibers network visible due to cracks in the cell tissue or to a lower cell density.

    Article Snippet: The murine mesenchymal stem cell line C3H10T1/2 [ ] (ATCC CCL-26) initially proliferated in culture flasks at 37 °C, 5% CO2 , in 1 g/L glucose DMEM medium (Sigma-Aldrich, St. Louis, MO, USA) complemented with 10% of Fetal Bovine Serum (Gibco Invitrogen, Waltham, MA USA), 2% of penicillin-streptomycin (Gibco Invitrogen, Waltham, MA, USA) and 2% of 200 mM l -glutamine (Gibco Invitrogen, Waltham, MA, USA).

    Techniques: Fluorescence, Microscopy, Cell Culture, Proximity Ligation Assay, Ethidium Homodimer Assay

    The effects of Sirt1 pharmacologic activation and inhibition on adipogenic markers in C3HT101/2 cells (A) . Oil-red-o staining in C3H10T1/2 cells induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (B,C) Gene expression analysis of adipocyte (B) and mitochondrial markers (C) induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (D) Gene expression analysis of adipocyte markers induced to adipogenesis and supplemented with Ex527 or vehicle (DMSO). Results are Mean ± SEM. * P

    Journal: Frontiers in Endocrinology

    Article Title: Sirt1 Promotes a Thermogenic Gene Program in Bone Marrow Adipocytes: From Mice to (Wo)Men

    doi: 10.3389/fendo.2019.00126

    Figure Lengend Snippet: The effects of Sirt1 pharmacologic activation and inhibition on adipogenic markers in C3HT101/2 cells (A) . Oil-red-o staining in C3H10T1/2 cells induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (B,C) Gene expression analysis of adipocyte (B) and mitochondrial markers (C) induced to adipogenesis and supplemented with SRT3025 or vehicle (DMSO). (D) Gene expression analysis of adipocyte markers induced to adipogenesis and supplemented with Ex527 or vehicle (DMSO). Results are Mean ± SEM. * P

    Article Snippet: Experiments in the Murine Mesenchymal Stem-Cell Line C3H10T1/2 The C3H10T1/2 (ATCC CCL-226) murine mesenchymal stem cell line is an established cell line model to investigate bone marrow adipocytes ( , ).

    Techniques: Activation Assay, Inhibition, Staining, Expressing

    The effect of Sirt1 over-expression on adipogenesis in C3HT101/2 cells (A) . Oil-red-o staining in Sirt1 over-expressing ( OE ) and C3H10T1/2 cells induced to adipogenesis. Data is presented as optical density (OD) corrected for cell number (crystal violet staining). Scale bar 500μm. (B,C) Immunoblot of Pgc1α and Prdm16 in Sirt1 OE and C3H10T1/2 cells 7 days post induction to adipogenesis. Results are Mean ± SEM. * P

    Journal: Frontiers in Endocrinology

    Article Title: Sirt1 Promotes a Thermogenic Gene Program in Bone Marrow Adipocytes: From Mice to (Wo)Men

    doi: 10.3389/fendo.2019.00126

    Figure Lengend Snippet: The effect of Sirt1 over-expression on adipogenesis in C3HT101/2 cells (A) . Oil-red-o staining in Sirt1 over-expressing ( OE ) and C3H10T1/2 cells induced to adipogenesis. Data is presented as optical density (OD) corrected for cell number (crystal violet staining). Scale bar 500μm. (B,C) Immunoblot of Pgc1α and Prdm16 in Sirt1 OE and C3H10T1/2 cells 7 days post induction to adipogenesis. Results are Mean ± SEM. * P

    Article Snippet: Experiments in the Murine Mesenchymal Stem-Cell Line C3H10T1/2 The C3H10T1/2 (ATCC CCL-226) murine mesenchymal stem cell line is an established cell line model to investigate bone marrow adipocytes ( , ).

    Techniques: Over Expression, Staining, Expressing