human β-actin Search Results


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  • 92
    Stratagene human β actin
    Cacna2d2 is predominantly expressed in brain in a pattern distinct from Cacna2d1 but similar to Cacna2d3 . a , Expression of Cacna2d2 , Cacna2d1 , and Cacna2d3 by RT-PCR of P28 +/+ mouse tissue RNA. <t>β-Actin</t> primers were used to allow comparisons of transcript levels. Negative control was no RNA. i , Cacna2d2 -12F/14R; ii , Cacna2d1 -1F/1R; iii , Cacna2d3 -1F/1R; iv , β-actin. b , In situ hybridization analyses of whole-brain sections (P21 +/+) with a DIG-labeled antisense Cacna2d2 RNA probe (nt 3705–4909). i , Sagittal section demonstrates the highest level of expression in cerebellum ( cb ), with some expression also in medulla ( m ), pons ( p ), and striatum ( st ). ii , The horizontal section shows expression in cortex ( cx ), nucleus reticularis thalami ( nrt ), habenula ( ha ), and hippocampus ( h ). c , Detailed examination of some of these areas by in situ hybridization with the same probe as above. Moderate expression is seen in medulla ( i ), and higher levels were seen in a subpopulation of cells of the striatum ( ii ) and cerebral cortex in a large proportion of cortical neurons throughout all layers ( iii ). Uniform expression is seen in nRT ( iv ), habenula ( v ), and hippocampus ( vi ). Scale bar: Ci–Ciii , 100 μ m; Civ , Cv , 50 μm.
    Human β Actin, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore human β actin
    MFAP-4 over-expression prevents UVB-induced deterioration of collagen I. (a) Immunohistochemical analysis with a rabbit anti-human collagen I antibody or a non-specific control rabbit IgG was carried out using paraffin embedded sections from the xenografted skin treated with a lentiviral vector encoding a control reporter gene with or without continuous UVB irradiation for 8 wks and using xenografted skin over-expressing MFAP-4 with UVB exposure for 8 wks. Scale bars = 100 μm. (b) Western blotting analysis with an anti-human collagen I or an anti-human <t>β-actin</t> antibody to assess the protein levels of collagen I in xenografted skin treated with lentiviral vectors encoding a control reporter gene or MFAP-4 before and after UVB exposure for 4 wks. (c) NHDFs were transfected with siRNAs for non-specific sequences (Control) or MFAP-4 twice during the culture for 8 days. Control siRNA-transfected cells were treated with or without 10 nM human recombinant MFAP-4 during the culture. Quantitative RT-PCR analysis of the MMP-1 mRNA expression in cultured NHDFs was performed as described in the legend of Fig. 5a . The values reported represent means ± SD. ***p
    Human β Actin, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human β actin
    Effect of transcutaneous application of CO 2 treatment on mitochondrial proliferation in tumors. qRT-PCR for PGC-1α (A) and TFAM (B) in CO 2 treated or control tumor specimens collected two weeks post-treatment. Expression was normalized to <t>β-actin</t> control. Data represent the mean ± S.E of at least three independent experiments (* p
    Human β Actin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 798 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SABiosciences human β actin
    Regional distribution of LRRK2 protein and miR-205 expression in mouse brains. ( A ) Western blot of LRRK2 protein in different brain regions of 1-month-old mice. CTX, cerebral cortex; HIP, hippocampus; MB, midbrain; STR, striatum; CB, cerebellum. TH and <t>β-actin</t> were used as loading controls. ( B–E ) Bar graphs quantify the expression of LRRK2 protein (B), mRNA (C), miR-205 (D) and miR-452 as a negative control (E) in different brain regions. The relative expression is shown as a percentage of the mean using the cortex region as reference. *** P
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    TaKaRa human β actin
    Fluorescence imaging of mTFP1 fusion constructs . (A)-(K) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mTFP1-α-actinin-19 (human non-muscle); (B) mTFP1-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mTFP1-Cx43-7 (rat α-1 connexin-43); (D) mTFP1-keratin-17 (human cytokeratin 18); (E) mTFP1-endoplasmic reticulum-3 (calreticulin signal sequence (51 nucleotides) and KDEL retention sequence); (F) mTFP1-paxillin-22 (chicken); (G) mTFP1-EB3-7 (human microtubule-associated protein; RP/EB family); (H) mTFP1-lysosomes-20 (rat lysosomal membrane glycoprotein 1); (I) mTFP1-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (J) mTFP1-vimentin-7 (human); (K) mTFP1-zyxin-7 (human). (L)-(T) C-terminal fusion constructs: (L) mTFP1-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (M) mTFP1-lamin B1–10 (human); (N) <t>mTFP1-β-actin-7;</t> (O) mTFP1-clathrin light chain-15 (human); (P) mTFP1-fibrillarin-7 (human); (Q) mTFP1-vinculin-23 (human); (R) mTFP1-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S)mTFP1-β-tubulin-6 (human); (T) mTFP1-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras). The cell line used for expressing mTFP1 fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (A), (G), (K), (N) and Q) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
    Human β Actin, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human β actin
    Fluorescence imaging of mTFP1 fusion constructs . (A)-(K) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mTFP1-α-actinin-19 (human non-muscle); (B) mTFP1-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mTFP1-Cx43-7 (rat α-1 connexin-43); (D) mTFP1-keratin-17 (human cytokeratin 18); (E) mTFP1-endoplasmic reticulum-3 (calreticulin signal sequence (51 nucleotides) and KDEL retention sequence); (F) mTFP1-paxillin-22 (chicken); (G) mTFP1-EB3-7 (human microtubule-associated protein; RP/EB family); (H) mTFP1-lysosomes-20 (rat lysosomal membrane glycoprotein 1); (I) mTFP1-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (J) mTFP1-vimentin-7 (human); (K) mTFP1-zyxin-7 (human). (L)-(T) C-terminal fusion constructs: (L) mTFP1-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (M) mTFP1-lamin B1–10 (human); (N) <t>mTFP1-β-actin-7;</t> (O) mTFP1-clathrin light chain-15 (human); (P) mTFP1-fibrillarin-7 (human); (Q) mTFP1-vinculin-23 (human); (R) mTFP1-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S)mTFP1-β-tubulin-6 (human); (T) mTFP1-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras). The cell line used for expressing mTFP1 fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (A), (G), (K), (N) and Q) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
    Human β Actin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology human β actin
    The protein levels of NF-κB in the BEAS-2B cells at passages 10, 20 and 30. ( A ) The protein bands of NF-κB in the BEAS-2B cells at passage 10, 20 and 30 in four groups were shown by Western blot. 1–4 bands: B(a)P, CTPE, DMSO, Co-culture+CTPE. ( B ) Quantitative comparison of the densitometry of NF-κB protein bands intensities normalized to the loading control <t>β-actin.</t> The level of the relative densitometry of NF-κB protein in Co-culture/CTPE group started to increase at passage 10, which was higher than that of CTPE or B(a)P at passage 20, and kept the highest level at passage 30.(n = 6, *: vs DMSO, P
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    Cytoskeleton Inc human β actin
    Monoclonal anti-ORF1p Δ1–143 antibodies (rG24) specifically recognize ORF1-encoded proteins. ( A ) Recombinant 33-kDa ORF1p Δ1–143 was used for the generation of monoclonal antibodies. MW, molecular weight marker. ( B ) Immunoblot detection of 10 ng of purified His-tagged ORF1p Δ1–143 with rG24 (lane 1) and anti-His-tag antibodies (lane 2). ( C ) Immunoblot analysis examining the specificity of the generated monoclonal anti-ORF1p antibody. Sixty microgram whole cell extracts from pORF1p Δ1–143 -IRESpuro (lane 2)- or pIRESpuro-transfected (lane 3) REF cells were loaded on a 12% SDS–PAA gel. In contrast to the parental empty expression vector pIRESpuro, pORF1p Δ1–143 -IRESpuro is expressing the N-terminally truncated 33-kDa L1Rn-ORF1 protein, which is detected by the rG24 antibody. As a loading control the membrane was stripped and incubated with an <t>anti-β-actin</t> antibody; lane 1, 10 ng purified ORF1p Δ1–143 protein; ( D – F ) Confocal images of REF cells transfected with ORF1p- (D, E) and ORF1p Δ1–143 - (F) expressing constructs. ORF1-encoded proteins and vimentin were stained using monoclonal rG24 (green) and polyclonal anti-vimentin antibodies (red), respectively. The co-localization channel (yellow) was generated using the ImarisColoc module. Cytoplasmic ORF1p aggregates co-localize with vimentin filaments partially. Scale bar—10 µm.
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    Rockland Immunochemicals human β actin
    Monoclonal anti-ORF1p Δ1–143 antibodies (rG24) specifically recognize ORF1-encoded proteins. ( A ) Recombinant 33-kDa ORF1p Δ1–143 was used for the generation of monoclonal antibodies. MW, molecular weight marker. ( B ) Immunoblot detection of 10 ng of purified His-tagged ORF1p Δ1–143 with rG24 (lane 1) and anti-His-tag antibodies (lane 2). ( C ) Immunoblot analysis examining the specificity of the generated monoclonal anti-ORF1p antibody. Sixty microgram whole cell extracts from pORF1p Δ1–143 -IRESpuro (lane 2)- or pIRESpuro-transfected (lane 3) REF cells were loaded on a 12% SDS–PAA gel. In contrast to the parental empty expression vector pIRESpuro, pORF1p Δ1–143 -IRESpuro is expressing the N-terminally truncated 33-kDa L1Rn-ORF1 protein, which is detected by the rG24 antibody. As a loading control the membrane was stripped and incubated with an <t>anti-β-actin</t> antibody; lane 1, 10 ng purified ORF1p Δ1–143 protein; ( D – F ) Confocal images of REF cells transfected with ORF1p- (D, E) and ORF1p Δ1–143 - (F) expressing constructs. ORF1-encoded proteins and vimentin were stained using monoclonal rG24 (green) and polyclonal anti-vimentin antibodies (red), respectively. The co-localization channel (yellow) was generated using the ImarisColoc module. Cytoplasmic ORF1p aggregates co-localize with vimentin filaments partially. Scale bar—10 µm.
    Human β Actin, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    US Biological Life Sciences human β actin
    ( a ) Effect of EMP2 on FAK phosphorylation. Levels of total FAK, p-FAK (Y397, Y407, Y861, and Y925), and <t>β</t> -actin were measured by Western blot analysis in the cell lines ARPE-19 and ARPE-19/EMP2 and were quantified by densitometry. ( b ) p-Src (Tyr
    Human β Actin, supplied by US Biological Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene human β actin
    ( a ) Effect of EMP2 on FAK phosphorylation. Levels of total FAK, p-FAK (Y397, Y407, Y861, and Y925), and <t>β</t> -actin were measured by Western blot analysis in the cell lines ARPE-19 and ARPE-19/EMP2 and were quantified by densitometry. ( b ) p-Src (Tyr
    Human β Actin, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Virostat human β actin
    Effects of bile acids on the expression of HCV NS5B in GS4.1 cells. Unconjugated bile acid CDCA or DCA was incubated in 1- or 2-day-old semiconfluent GS4.1 cells for 48 h. Cell lysates were prepared after the incubation, and Western blot analysis was performed for the expression of HCV NS5B. As a loading control, Western blot analysis of <t>β-actin</t> was performed with the same samples.
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    Cell Signaling Technology Inc human β actin
    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and <t>β-actin.</t>
    Human β Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human β actin amplicon
    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and <t>β-actin.</t>
    Human β Actin Amplicon, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory human β actin promoter
    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and <t>β-actin.</t>
    Human β Actin Promoter, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 89/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene human β actin probe
    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and <t>β-actin.</t>
    Human β Actin Probe, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa human β actin primer
    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and <t>β-actin.</t>
    Human β Actin Primer, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam human β actin
    SMAD3 Mediates Both the BTIC-Promoting and BTIC-Suppressing Programs of TGF-β (A) MDA-MB-231 and HCC-1954 first generation mammosphere cultures with and without addition of TGF-β. TGF-β was added to the media at the moment of cell seeding and mammospheres were allowed to form for 7 days. Note that this is not a quantitative assay. (B) Western blot showing SMAD2 phosphorylation levels upon TGF-β pathway induction. 7-day-old mammospheres were treated with exogenous TGF-β ligand for 1 hr. Total SMAD2/3 and <t>β-actin</t> levels were used as loading controls. (C) Comparison of TGF-β-dependent genes in MDA-MB-231 and HCC-1954 BTICs. MDA-MB-231 and HCC-1954 cells were grown as mammospheres for 7 days and treated with TGF-β for 1, 3, 6, and 24 hr, and gene expression profiling was performed using Illumina HumanHT-12 BeadChips. The plot shows the number of TGF-β-dependent genes in each BTIC at each time point (false discovery rate [FDR]
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    TaKaRa human β actin probe
    Expression of Elm1 and Cyr61 after serum stimulation of BALB/c 3T3 cells. 2 μg of poly(A) + RNA resolved electrophoretically were hybridized with the Elm1 ( A ) or Cyr61 ( B ) cDNA probe. The membrane was rehybridized with a human <t>β-actin</t> probe ( C ). poly(A) + RNA from quiescent cells (lane 1 ), cells stimulated with serum for 30 min (lane 2 ), and 3 h (lane 3 ) were loaded.
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    FUJIFILM human β actin probe
    Csk expression and the kinase activity of synoviocytes. ( a ) Western blot analysis shows that Csk virus induced Csk expression in human synovial cells in an moi-dependent manner without significant changes in expression of c-Src protein. Fold increase in Csk expression normalized by <t>β-actin</t> is shown below each blot. The results shown are from a single experiment typical of 3 experiments yielding identical results. ( b ) Immunofluorescence staining using anti-Csk antibody shows that WT virus–infected synoviocytes expressed a very small amount of Csk protein. ( c ) Diffuse cytoplasmic staining in Csk virus–infected (moi = 100) synoviocytes. ( d ) In vitro kinase assay, using enolase as a substrate demonstrated that kinase activity of c-Src, Fyn, and c-Yes was remarkably repressed in Csk-overexpressing synoviocytes, but the expression of each protein was unaffected by Csk expression. Relative kinase activity of each kinase is shown below. The results were derived from 3 separate experiments. Suppression on each kinase was statistically significant (* P
    Human β Actin Probe, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human β actin primers
    Representative semiquantitative RT-PCR gel image demonstrating expression of testicular genes in calorie restricted ( CR ; n = 5) or control ( CON ; n = 5) young adult (∼12 years old) rhesus macaques. The housekeeping gene <t>β-actin</t> was used as a positive control and for normalizing images for analysis. No significant differences (mean ± SEM; P > 0.05) in gene expression were observed between the treatment groups
    Human β Actin Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioNex Solutions human β actin primers
    Effect of chloroform fraction from Carpinus tschonoskii on the STAT1 signal in IFN-γ-stimulated HaCaT human keratinocytes. (A) Cells were pretreated with the indicated concentration of chloroform fraction of C. tschonoskii for 2 hr, and then the phosphorylation of STAT1 was determined in cells stimulated by IFN-γ (10 ng/ml) for 30 min. The levels of STAT1 and <t>β-actin</t> were identified using Western blotting analysis. (B) Cells were pretreated with the chloroform fraction (50 μg/ml) of C. tschonoskii for 2 hr, and then the translocation of STAT1 protein was determined in cells stimulated by IFN-γ (10 ng/ml) for 60 min. Immunofluorescence stain of STAT1 was stained with DyLight488-conjugated 2 nd antibody and the fluorescence was identified using confocal microscopy (FV500, OLYMPUS) and the images were acquired at constant conditions (PMT, gain, offset, magnification (20 × objective with zoom factor of 2.5) and resolution, etc.). These data are representative of three independent experiments.
    Human β Actin Primers, supplied by BioNex Solutions, used in various techniques. Bioz Stars score: 88/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson human β actin primers
    Effect of chloroform fraction from Carpinus tschonoskii on the STAT1 signal in IFN-γ-stimulated HaCaT human keratinocytes. (A) Cells were pretreated with the indicated concentration of chloroform fraction of C. tschonoskii for 2 hr, and then the phosphorylation of STAT1 was determined in cells stimulated by IFN-γ (10 ng/ml) for 30 min. The levels of STAT1 and <t>β-actin</t> were identified using Western blotting analysis. (B) Cells were pretreated with the chloroform fraction (50 μg/ml) of C. tschonoskii for 2 hr, and then the translocation of STAT1 protein was determined in cells stimulated by IFN-γ (10 ng/ml) for 60 min. Immunofluorescence stain of STAT1 was stained with DyLight488-conjugated 2 nd antibody and the fluorescence was identified using confocal microscopy (FV500, OLYMPUS) and the images were acquired at constant conditions (PMT, gain, offset, magnification (20 × objective with zoom factor of 2.5) and resolution, etc.). These data are representative of three independent experiments.
    Human β Actin Primers, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human β actin primers/product/Becton Dickinson
    Average 88 stars, based on 8 article reviews
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    human β actin primers - by Bioz Stars, 2020-08
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    91
    OriGene human β actin control primers
    Effect of chloroform fraction from Carpinus tschonoskii on the STAT1 signal in IFN-γ-stimulated HaCaT human keratinocytes. (A) Cells were pretreated with the indicated concentration of chloroform fraction of C. tschonoskii for 2 hr, and then the phosphorylation of STAT1 was determined in cells stimulated by IFN-γ (10 ng/ml) for 30 min. The levels of STAT1 and <t>β-actin</t> were identified using Western blotting analysis. (B) Cells were pretreated with the chloroform fraction (50 μg/ml) of C. tschonoskii for 2 hr, and then the translocation of STAT1 protein was determined in cells stimulated by IFN-γ (10 ng/ml) for 60 min. Immunofluorescence stain of STAT1 was stained with DyLight488-conjugated 2 nd antibody and the fluorescence was identified using confocal microscopy (FV500, OLYMPUS) and the images were acquired at constant conditions (PMT, gain, offset, magnification (20 × objective with zoom factor of 2.5) and resolution, etc.). These data are representative of three independent experiments.
    Human β Actin Control Primers, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human β actin control primers/product/OriGene
    Average 91 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    human β actin control primers - by Bioz Stars, 2020-08
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    Image Search Results


    Cacna2d2 is predominantly expressed in brain in a pattern distinct from Cacna2d1 but similar to Cacna2d3 . a , Expression of Cacna2d2 , Cacna2d1 , and Cacna2d3 by RT-PCR of P28 +/+ mouse tissue RNA. β-Actin primers were used to allow comparisons of transcript levels. Negative control was no RNA. i , Cacna2d2 -12F/14R; ii , Cacna2d1 -1F/1R; iii , Cacna2d3 -1F/1R; iv , β-actin. b , In situ hybridization analyses of whole-brain sections (P21 +/+) with a DIG-labeled antisense Cacna2d2 RNA probe (nt 3705–4909). i , Sagittal section demonstrates the highest level of expression in cerebellum ( cb ), with some expression also in medulla ( m ), pons ( p ), and striatum ( st ). ii , The horizontal section shows expression in cortex ( cx ), nucleus reticularis thalami ( nrt ), habenula ( ha ), and hippocampus ( h ). c , Detailed examination of some of these areas by in situ hybridization with the same probe as above. Moderate expression is seen in medulla ( i ), and higher levels were seen in a subpopulation of cells of the striatum ( ii ) and cerebral cortex in a large proportion of cortical neurons throughout all layers ( iii ). Uniform expression is seen in nRT ( iv ), habenula ( v ), and hippocampus ( vi ). Scale bar: Ci–Ciii , 100 μ m; Civ , Cv , 50 μm.

    Journal: The Journal of Neuroscience

    Article Title: Ducky Mouse Phenotype of Epilepsy and Ataxia Is Associated with Mutations in the Cacna2d2 Gene and Decreased Calcium Channel Current in Cerebellar Purkinje Cells

    doi: 10.1523/JNEUROSCI.21-16-06095.2001

    Figure Lengend Snippet: Cacna2d2 is predominantly expressed in brain in a pattern distinct from Cacna2d1 but similar to Cacna2d3 . a , Expression of Cacna2d2 , Cacna2d1 , and Cacna2d3 by RT-PCR of P28 +/+ mouse tissue RNA. β-Actin primers were used to allow comparisons of transcript levels. Negative control was no RNA. i , Cacna2d2 -12F/14R; ii , Cacna2d1 -1F/1R; iii , Cacna2d3 -1F/1R; iv , β-actin. b , In situ hybridization analyses of whole-brain sections (P21 +/+) with a DIG-labeled antisense Cacna2d2 RNA probe (nt 3705–4909). i , Sagittal section demonstrates the highest level of expression in cerebellum ( cb ), with some expression also in medulla ( m ), pons ( p ), and striatum ( st ). ii , The horizontal section shows expression in cortex ( cx ), nucleus reticularis thalami ( nrt ), habenula ( ha ), and hippocampus ( h ). c , Detailed examination of some of these areas by in situ hybridization with the same probe as above. Moderate expression is seen in medulla ( i ), and higher levels were seen in a subpopulation of cells of the striatum ( ii ) and cerebral cortex in a large proportion of cortical neurons throughout all layers ( iii ). Uniform expression is seen in nRT ( iv ), habenula ( v ), and hippocampus ( vi ). Scale bar: Ci–Ciii , 100 μ m; Civ , Cv , 50 μm.

    Article Snippet: Northern blot analysis of 10 μg of cerebellar mRNA using Duralon UV nylon membrane and full-length Cacna2d2 or human β actin as probes (Stratagene, La Jolla, CA) was performed using the suggested conditions of the manufacturer to optimize the identification of the wild-type 5.5 kb Cacna2d2 transcript.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Negative Control, In Situ Hybridization, Labeling

    The du mutation is a genomic rearrangement involving the Cacna2d2 gene. a , Two mutant transcripts can be identified by RT-PCR of total brain RNA from du/du mice. Two +/+, two du/du samples, and a negative control (no RNA) are shown per gel. i , Top , Normal size amplification product of exons 1–3 is shown in +/+ and du/du RNA, with reduced levels in the latter. Middle , Amplification between exons 1 and 4 does not produce a product in du/du RNA, suggesting disruption of the Cacna2d2 gene in this region. Bottom , Amplification of the du- specific chimeric transcript of Cacna2d2 exons 1, 2, and 3 and a novel sequence X. ii , Overlapping PCR fragments spanning exons 2–39 of Cacna2d2 can be detected in +/+ and du/du RNA, with lower levels observed in du/du samples. b , Wild-type Cacna2d2 transcript (5.5 kb) is absent from du/du brain by Northern analysis using cerebellar mRNA and full-length Cacna2d2 as a probe. Low levels of two du -specific bands (∼1.5 and 5 kb) are detected. The filter was rehybridized with β-actin as a control for RNA loading. c , PFGE shows duplication of Cacna2d2 exons 2 and 3 and region X in du/du genomic DNA. Southern analysis of Not I-digested genomic DNA separated by PFGE from +/+, +/ du , and du/du mice is shown. Blots were hybridized with Cacna2d2 probes: i , exon 1; ii , exons 2–3; iii , exons 4–39; iv , region X. Sizes are in kilobases. d , A scale representation of the genomic region containing Cacna2d2 ( red ) and region X ( blue ) in +/+ and du/du mice. N , Not I sites. The presence of one or two B2 repeats 5′ to region X is marked by a vertical line . The mutant transcripts 1 and 2 produced from each region in du/du are represented by colored boxes . The Cacna2d2 gene is arranged 5′ to 3′ in +/+. In du/du , exons encoding mutant transcript 1 are shown in a 5′ to 3′ direction, and those encoding mutant transcript 2 are inverted and shown 3′ to 5′. The distance between exon 3 and region X is unknown but is > 12 kb. The scale bar is in kilobases.

    Journal: The Journal of Neuroscience

    Article Title: Ducky Mouse Phenotype of Epilepsy and Ataxia Is Associated with Mutations in the Cacna2d2 Gene and Decreased Calcium Channel Current in Cerebellar Purkinje Cells

    doi: 10.1523/JNEUROSCI.21-16-06095.2001

    Figure Lengend Snippet: The du mutation is a genomic rearrangement involving the Cacna2d2 gene. a , Two mutant transcripts can be identified by RT-PCR of total brain RNA from du/du mice. Two +/+, two du/du samples, and a negative control (no RNA) are shown per gel. i , Top , Normal size amplification product of exons 1–3 is shown in +/+ and du/du RNA, with reduced levels in the latter. Middle , Amplification between exons 1 and 4 does not produce a product in du/du RNA, suggesting disruption of the Cacna2d2 gene in this region. Bottom , Amplification of the du- specific chimeric transcript of Cacna2d2 exons 1, 2, and 3 and a novel sequence X. ii , Overlapping PCR fragments spanning exons 2–39 of Cacna2d2 can be detected in +/+ and du/du RNA, with lower levels observed in du/du samples. b , Wild-type Cacna2d2 transcript (5.5 kb) is absent from du/du brain by Northern analysis using cerebellar mRNA and full-length Cacna2d2 as a probe. Low levels of two du -specific bands (∼1.5 and 5 kb) are detected. The filter was rehybridized with β-actin as a control for RNA loading. c , PFGE shows duplication of Cacna2d2 exons 2 and 3 and region X in du/du genomic DNA. Southern analysis of Not I-digested genomic DNA separated by PFGE from +/+, +/ du , and du/du mice is shown. Blots were hybridized with Cacna2d2 probes: i , exon 1; ii , exons 2–3; iii , exons 4–39; iv , region X. Sizes are in kilobases. d , A scale representation of the genomic region containing Cacna2d2 ( red ) and region X ( blue ) in +/+ and du/du mice. N , Not I sites. The presence of one or two B2 repeats 5′ to region X is marked by a vertical line . The mutant transcripts 1 and 2 produced from each region in du/du are represented by colored boxes . The Cacna2d2 gene is arranged 5′ to 3′ in +/+. In du/du , exons encoding mutant transcript 1 are shown in a 5′ to 3′ direction, and those encoding mutant transcript 2 are inverted and shown 3′ to 5′. The distance between exon 3 and region X is unknown but is > 12 kb. The scale bar is in kilobases.

    Article Snippet: Northern blot analysis of 10 μg of cerebellar mRNA using Duralon UV nylon membrane and full-length Cacna2d2 or human β actin as probes (Stratagene, La Jolla, CA) was performed using the suggested conditions of the manufacturer to optimize the identification of the wild-type 5.5 kb Cacna2d2 transcript.

    Techniques: Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Negative Control, Amplification, Sequencing, Polymerase Chain Reaction, Northern Blot, Produced

    MFAP-4 over-expression prevents UVB-induced deterioration of collagen I. (a) Immunohistochemical analysis with a rabbit anti-human collagen I antibody or a non-specific control rabbit IgG was carried out using paraffin embedded sections from the xenografted skin treated with a lentiviral vector encoding a control reporter gene with or without continuous UVB irradiation for 8 wks and using xenografted skin over-expressing MFAP-4 with UVB exposure for 8 wks. Scale bars = 100 μm. (b) Western blotting analysis with an anti-human collagen I or an anti-human β-actin antibody to assess the protein levels of collagen I in xenografted skin treated with lentiviral vectors encoding a control reporter gene or MFAP-4 before and after UVB exposure for 4 wks. (c) NHDFs were transfected with siRNAs for non-specific sequences (Control) or MFAP-4 twice during the culture for 8 days. Control siRNA-transfected cells were treated with or without 10 nM human recombinant MFAP-4 during the culture. Quantitative RT-PCR analysis of the MMP-1 mRNA expression in cultured NHDFs was performed as described in the legend of Fig. 5a . The values reported represent means ± SD. ***p

    Journal: Scientific Reports

    Article Title: Essential role of microfibrillar-associated protein 4 in human cutaneous homeostasis and in its photoprotection

    doi: 10.1038/srep00164

    Figure Lengend Snippet: MFAP-4 over-expression prevents UVB-induced deterioration of collagen I. (a) Immunohistochemical analysis with a rabbit anti-human collagen I antibody or a non-specific control rabbit IgG was carried out using paraffin embedded sections from the xenografted skin treated with a lentiviral vector encoding a control reporter gene with or without continuous UVB irradiation for 8 wks and using xenografted skin over-expressing MFAP-4 with UVB exposure for 8 wks. Scale bars = 100 μm. (b) Western blotting analysis with an anti-human collagen I or an anti-human β-actin antibody to assess the protein levels of collagen I in xenografted skin treated with lentiviral vectors encoding a control reporter gene or MFAP-4 before and after UVB exposure for 4 wks. (c) NHDFs were transfected with siRNAs for non-specific sequences (Control) or MFAP-4 twice during the culture for 8 days. Control siRNA-transfected cells were treated with or without 10 nM human recombinant MFAP-4 during the culture. Quantitative RT-PCR analysis of the MMP-1 mRNA expression in cultured NHDFs was performed as described in the legend of Fig. 5a . The values reported represent means ± SD. ***p

    Article Snippet: Samples were transferred to PVDF membranes (Bio-Rad Laboratories) and were incubated with antibodies specific for human MFAP-4 (AdipoGen, Incheon, Korea), human collagen I (United States Biological, Swampscott, MA), human MMP-1 (AbD Serotec, Oxford, UK), human MMP-12 (Millipore), human fibrillin-1 (Elastin Products Company) or human β-actin (Sigma).

    Techniques: Over Expression, Immunohistochemistry, Plasmid Preparation, Irradiation, Expressing, Western Blot, Transfection, Recombinant, Quantitative RT-PCR, Cell Culture

    Effect of transcutaneous application of CO 2 treatment on mitochondrial proliferation in tumors. qRT-PCR for PGC-1α (A) and TFAM (B) in CO 2 treated or control tumor specimens collected two weeks post-treatment. Expression was normalized to β-actin control. Data represent the mean ± S.E of at least three independent experiments (* p

    Journal: PLoS ONE

    Article Title: Transcutaneous Application of Carbon Dioxide (CO2) Induces Mitochondrial Apoptosis in Human Malignant Fibrous Histiocytoma In Vivo

    doi: 10.1371/journal.pone.0049189

    Figure Lengend Snippet: Effect of transcutaneous application of CO 2 treatment on mitochondrial proliferation in tumors. qRT-PCR for PGC-1α (A) and TFAM (B) in CO 2 treated or control tumor specimens collected two weeks post-treatment. Expression was normalized to β-actin control. Data represent the mean ± S.E of at least three independent experiments (* p

    Article Snippet: Pre-designed primers specific for human PGC-1α , human TFAM and human β-actin were obtained from Invitrogen (Carlsbad, CA, USA).

    Techniques: Quantitative RT-PCR, Pyrolysis Gas Chromatography, Expressing

    Regional distribution of LRRK2 protein and miR-205 expression in mouse brains. ( A ) Western blot of LRRK2 protein in different brain regions of 1-month-old mice. CTX, cerebral cortex; HIP, hippocampus; MB, midbrain; STR, striatum; CB, cerebellum. TH and β-actin were used as loading controls. ( B–E ) Bar graphs quantify the expression of LRRK2 protein (B), mRNA (C), miR-205 (D) and miR-452 as a negative control (E) in different brain regions. The relative expression is shown as a percentage of the mean using the cortex region as reference. *** P

    Journal: Human Molecular Genetics

    Article Title: MicroRNA-205 regulates the expression of Parkinson's disease-related leucine-rich repeat kinase 2 protein

    doi: 10.1093/hmg/dds470

    Figure Lengend Snippet: Regional distribution of LRRK2 protein and miR-205 expression in mouse brains. ( A ) Western blot of LRRK2 protein in different brain regions of 1-month-old mice. CTX, cerebral cortex; HIP, hippocampus; MB, midbrain; STR, striatum; CB, cerebellum. TH and β-actin were used as loading controls. ( B–E ) Bar graphs quantify the expression of LRRK2 protein (B), mRNA (C), miR-205 (D) and miR-452 as a negative control (E) in different brain regions. The relative expression is shown as a percentage of the mean using the cortex region as reference. *** P

    Article Snippet: The primers for the human β-actin were purchased from SABiosciences.

    Techniques: Expressing, Western Blot, Mouse Assay, Negative Control

    MiR-205 modulates LRRK2 protein expression in vitro . ( A , B ) Western blots show LRRK2 protein expression levels in pre-miR-205 oligo- or LRRK2 siRNA-transfected HEK293T cells (A) and primary cultured mouse cortical neurons (B). A scrambled microRNA oligo (Ctrl oligo), a scrambled siRNA (Ctrl siRNA) and a vehicle (Vcl) only were used as negative controls. β-Actin or β-III tubulin was used as a loading control. Bar graphs depict the signal intensities of LRRK2-positive bands in the vehicle, microRNA and siRNA-treated samples. ** P

    Journal: Human Molecular Genetics

    Article Title: MicroRNA-205 regulates the expression of Parkinson's disease-related leucine-rich repeat kinase 2 protein

    doi: 10.1093/hmg/dds470

    Figure Lengend Snippet: MiR-205 modulates LRRK2 protein expression in vitro . ( A , B ) Western blots show LRRK2 protein expression levels in pre-miR-205 oligo- or LRRK2 siRNA-transfected HEK293T cells (A) and primary cultured mouse cortical neurons (B). A scrambled microRNA oligo (Ctrl oligo), a scrambled siRNA (Ctrl siRNA) and a vehicle (Vcl) only were used as negative controls. β-Actin or β-III tubulin was used as a loading control. Bar graphs depict the signal intensities of LRRK2-positive bands in the vehicle, microRNA and siRNA-treated samples. ** P

    Article Snippet: The primers for the human β-actin were purchased from SABiosciences.

    Techniques: Expressing, In Vitro, Western Blot, Transfection, Cell Culture

    Fluorescence imaging of mTFP1 fusion constructs . (A)-(K) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mTFP1-α-actinin-19 (human non-muscle); (B) mTFP1-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mTFP1-Cx43-7 (rat α-1 connexin-43); (D) mTFP1-keratin-17 (human cytokeratin 18); (E) mTFP1-endoplasmic reticulum-3 (calreticulin signal sequence (51 nucleotides) and KDEL retention sequence); (F) mTFP1-paxillin-22 (chicken); (G) mTFP1-EB3-7 (human microtubule-associated protein; RP/EB family); (H) mTFP1-lysosomes-20 (rat lysosomal membrane glycoprotein 1); (I) mTFP1-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (J) mTFP1-vimentin-7 (human); (K) mTFP1-zyxin-7 (human). (L)-(T) C-terminal fusion constructs: (L) mTFP1-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (M) mTFP1-lamin B1–10 (human); (N) mTFP1-β-actin-7; (O) mTFP1-clathrin light chain-15 (human); (P) mTFP1-fibrillarin-7 (human); (Q) mTFP1-vinculin-23 (human); (R) mTFP1-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S)mTFP1-β-tubulin-6 (human); (T) mTFP1-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras). The cell line used for expressing mTFP1 fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (A), (G), (K), (N) and Q) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

    Journal: BMC Biology

    Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

    doi: 10.1186/1741-7007-6-13

    Figure Lengend Snippet: Fluorescence imaging of mTFP1 fusion constructs . (A)-(K) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mTFP1-α-actinin-19 (human non-muscle); (B) mTFP1-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mTFP1-Cx43-7 (rat α-1 connexin-43); (D) mTFP1-keratin-17 (human cytokeratin 18); (E) mTFP1-endoplasmic reticulum-3 (calreticulin signal sequence (51 nucleotides) and KDEL retention sequence); (F) mTFP1-paxillin-22 (chicken); (G) mTFP1-EB3-7 (human microtubule-associated protein; RP/EB family); (H) mTFP1-lysosomes-20 (rat lysosomal membrane glycoprotein 1); (I) mTFP1-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (J) mTFP1-vimentin-7 (human); (K) mTFP1-zyxin-7 (human). (L)-(T) C-terminal fusion constructs: (L) mTFP1-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (M) mTFP1-lamin B1–10 (human); (N) mTFP1-β-actin-7; (O) mTFP1-clathrin light chain-15 (human); (P) mTFP1-fibrillarin-7 (human); (Q) mTFP1-vinculin-23 (human); (R) mTFP1-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S)mTFP1-β-tubulin-6 (human); (T) mTFP1-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras). The cell line used for expressing mTFP1 fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (A), (G), (K), (N) and Q) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

    Article Snippet: To prepare mTFP1 and mWasabi C-terminal fusions, the following digests were performed: human β-actin, NheI and BglII (Clontech); human α-tubulin, NheI and BglII (Clontech); human light chain clathrin, NheI and BglII (George Patterson, NIH); human lamin B1, NheI and BglII (George Patterson, NIH); human fibrillarin, AgeI and BglII (Evrogen); human vinculin, NheI and EcoRI (Open Biosystems, Huntsville, AL); peroximal targeting signal 1 (PTS1 – peroxisomes), AgeI and BspEI (Clontech); chicken protein tyrosine kinase 2, AgeI and BglII (Clare Waterman-Storer, NIH); human annexin (A4), AgeI and BspEI (Alen Piljic, EMBL, Heidelberg); human RhoB GTPase with an N-terminal c-Myc epitope tag (endosomes), AgeI and BspEI (Clontech); and the 20-amino acid farnesylation signal from c-Ha-Ras, AgeI and BspEI (membrane, Clontech).

    Techniques: Fluorescence, Imaging, Construct, Sequencing, Expressing

    Live cell imaging of mWasabi fusion vectors . (A)-(J) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

    Journal: BMC Biology

    Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

    doi: 10.1186/1741-7007-6-13

    Figure Lengend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

    Article Snippet: To prepare mTFP1 and mWasabi C-terminal fusions, the following digests were performed: human β-actin, NheI and BglII (Clontech); human α-tubulin, NheI and BglII (Clontech); human light chain clathrin, NheI and BglII (George Patterson, NIH); human lamin B1, NheI and BglII (George Patterson, NIH); human fibrillarin, AgeI and BglII (Evrogen); human vinculin, NheI and EcoRI (Open Biosystems, Huntsville, AL); peroximal targeting signal 1 (PTS1 – peroxisomes), AgeI and BspEI (Clontech); chicken protein tyrosine kinase 2, AgeI and BglII (Clare Waterman-Storer, NIH); human annexin (A4), AgeI and BspEI (Alen Piljic, EMBL, Heidelberg); human RhoB GTPase with an N-terminal c-Myc epitope tag (endosomes), AgeI and BspEI (Clontech); and the 20-amino acid farnesylation signal from c-Ha-Ras, AgeI and BspEI (membrane, Clontech).

    Techniques: Live Cell Imaging, Construct, Expressing

    The protein levels of NF-κB in the BEAS-2B cells at passages 10, 20 and 30. ( A ) The protein bands of NF-κB in the BEAS-2B cells at passage 10, 20 and 30 in four groups were shown by Western blot. 1–4 bands: B(a)P, CTPE, DMSO, Co-culture+CTPE. ( B ) Quantitative comparison of the densitometry of NF-κB protein bands intensities normalized to the loading control β-actin. The level of the relative densitometry of NF-κB protein in Co-culture/CTPE group started to increase at passage 10, which was higher than that of CTPE or B(a)P at passage 20, and kept the highest level at passage 30.(n = 6, *: vs DMSO, P

    Journal: PLoS ONE

    Article Title: Macrophages Facilitate Coal Tar Pitch Extract-Induced Tumorigenic Transformation of Human Bronchial Epithelial Cells Mediated by NF-?B

    doi: 10.1371/journal.pone.0051690

    Figure Lengend Snippet: The protein levels of NF-κB in the BEAS-2B cells at passages 10, 20 and 30. ( A ) The protein bands of NF-κB in the BEAS-2B cells at passage 10, 20 and 30 in four groups were shown by Western blot. 1–4 bands: B(a)P, CTPE, DMSO, Co-culture+CTPE. ( B ) Quantitative comparison of the densitometry of NF-κB protein bands intensities normalized to the loading control β-actin. The level of the relative densitometry of NF-κB protein in Co-culture/CTPE group started to increase at passage 10, which was higher than that of CTPE or B(a)P at passage 20, and kept the highest level at passage 30.(n = 6, *: vs DMSO, P

    Article Snippet: Immunoblots were blocked with 5% milk in TBS/0.1% Tween 20 for 1 h at room temperature, then incubated overnight at 4°C with 1∶500 rabbit polyclonal antibody against human NF-κB p65 (Santa Cruz) and 1∶1000 rabbit polyclonal antibody to human β-actin(Cell Singaling) in 3% milk/TBS/0.1% Tween 20, 1∶2000 goat anti-rabbit IgG (Santa Cruz Biotechnology) as secondary antibody in 3% milk/TBS/0.1% Tween 20 was used to incubate membranes at room temperature for 2 h. Proteins were detected by DAB detection system (Zhongshan Golden Bridge Bio.

    Techniques: Western Blot, Co-Culture Assay

    Monoclonal anti-ORF1p Δ1–143 antibodies (rG24) specifically recognize ORF1-encoded proteins. ( A ) Recombinant 33-kDa ORF1p Δ1–143 was used for the generation of monoclonal antibodies. MW, molecular weight marker. ( B ) Immunoblot detection of 10 ng of purified His-tagged ORF1p Δ1–143 with rG24 (lane 1) and anti-His-tag antibodies (lane 2). ( C ) Immunoblot analysis examining the specificity of the generated monoclonal anti-ORF1p antibody. Sixty microgram whole cell extracts from pORF1p Δ1–143 -IRESpuro (lane 2)- or pIRESpuro-transfected (lane 3) REF cells were loaded on a 12% SDS–PAA gel. In contrast to the parental empty expression vector pIRESpuro, pORF1p Δ1–143 -IRESpuro is expressing the N-terminally truncated 33-kDa L1Rn-ORF1 protein, which is detected by the rG24 antibody. As a loading control the membrane was stripped and incubated with an anti-β-actin antibody; lane 1, 10 ng purified ORF1p Δ1–143 protein; ( D – F ) Confocal images of REF cells transfected with ORF1p- (D, E) and ORF1p Δ1–143 - (F) expressing constructs. ORF1-encoded proteins and vimentin were stained using monoclonal rG24 (green) and polyclonal anti-vimentin antibodies (red), respectively. The co-localization channel (yellow) was generated using the ImarisColoc module. Cytoplasmic ORF1p aggregates co-localize with vimentin filaments partially. Scale bar—10 µm.

    Journal: Nucleic Acids Research

    Article Title: Functional endogenous LINE-1 retrotransposons are expressed and mobilized in rat chloroleukemia cells

    doi: 10.1093/nar/gkm1045

    Figure Lengend Snippet: Monoclonal anti-ORF1p Δ1–143 antibodies (rG24) specifically recognize ORF1-encoded proteins. ( A ) Recombinant 33-kDa ORF1p Δ1–143 was used for the generation of monoclonal antibodies. MW, molecular weight marker. ( B ) Immunoblot detection of 10 ng of purified His-tagged ORF1p Δ1–143 with rG24 (lane 1) and anti-His-tag antibodies (lane 2). ( C ) Immunoblot analysis examining the specificity of the generated monoclonal anti-ORF1p antibody. Sixty microgram whole cell extracts from pORF1p Δ1–143 -IRESpuro (lane 2)- or pIRESpuro-transfected (lane 3) REF cells were loaded on a 12% SDS–PAA gel. In contrast to the parental empty expression vector pIRESpuro, pORF1p Δ1–143 -IRESpuro is expressing the N-terminally truncated 33-kDa L1Rn-ORF1 protein, which is detected by the rG24 antibody. As a loading control the membrane was stripped and incubated with an anti-β-actin antibody; lane 1, 10 ng purified ORF1p Δ1–143 protein; ( D – F ) Confocal images of REF cells transfected with ORF1p- (D, E) and ORF1p Δ1–143 - (F) expressing constructs. ORF1-encoded proteins and vimentin were stained using monoclonal rG24 (green) and polyclonal anti-vimentin antibodies (red), respectively. The co-localization channel (yellow) was generated using the ImarisColoc module. Cytoplasmic ORF1p aggregates co-localize with vimentin filaments partially. Scale bar—10 µm.

    Article Snippet: Bovine α-tubulin and human β-actin were purchased from Cytoskeleton, Inc. (Denver, CO, USA).

    Techniques: Recombinant, Molecular Weight, Marker, Purification, Generated, Transfection, Expressing, Plasmid Preparation, Incubation, Construct, Staining

    L1Rn ORF1p is able to form filaments that co-align with vimentin filaments. ( A and B ) In situ overlay of recombinant L1-21 ORF1p onto PFA-fixed and Triton X-100-extracted MEFs. Immunofluorescence images of L1Rn ORF1p (A) and vimentin (B) are shown in green and red colors, respectively. Each panel represents the 3D reconstruction of xy confocal sections. ( C ) ORF1p and vimentin filaments co-align partially. The co-localization channel (yellow) was generated using the ImarisColoc module. Scale bar—10 µm. ( D ) Dot blot overlay assay demonstrating interaction of ORF1p with vimentin. ORF1p was immobilized onto a nitrocellulose membrane (dots I–IV). Each ORF1p dot contained 2 μg of protein. After blocking, immobilized ORF1p was overlaid with an excess of soluble vimentin, α-tubulin or β-actin, respectively. Incubation was carried out for 60 min. In order to control for successful immobilization and immunoblotting procedure, 50 ng of soluble vimentin, α-tubulin and β-actin were immobilized on the nitrocellulose membrane in parallel (dots V–VII). Membranes were washed subsequently and submitted to immunoblot analyses with anti-vimentin, anti-α-tubulin or anti-β-actin antibodies, respectively, due to the schematic shown. In contrast to α-tubulin and β-actin, only vimentin can be detected with its specific antibody on the nitrocellulose membrane, after overlaying ORF1p dots with the respective IF protein (dots I–III). This is indicating that only vimentin is binding to ORF1p.

    Journal: Nucleic Acids Research

    Article Title: Functional endogenous LINE-1 retrotransposons are expressed and mobilized in rat chloroleukemia cells

    doi: 10.1093/nar/gkm1045

    Figure Lengend Snippet: L1Rn ORF1p is able to form filaments that co-align with vimentin filaments. ( A and B ) In situ overlay of recombinant L1-21 ORF1p onto PFA-fixed and Triton X-100-extracted MEFs. Immunofluorescence images of L1Rn ORF1p (A) and vimentin (B) are shown in green and red colors, respectively. Each panel represents the 3D reconstruction of xy confocal sections. ( C ) ORF1p and vimentin filaments co-align partially. The co-localization channel (yellow) was generated using the ImarisColoc module. Scale bar—10 µm. ( D ) Dot blot overlay assay demonstrating interaction of ORF1p with vimentin. ORF1p was immobilized onto a nitrocellulose membrane (dots I–IV). Each ORF1p dot contained 2 μg of protein. After blocking, immobilized ORF1p was overlaid with an excess of soluble vimentin, α-tubulin or β-actin, respectively. Incubation was carried out for 60 min. In order to control for successful immobilization and immunoblotting procedure, 50 ng of soluble vimentin, α-tubulin and β-actin were immobilized on the nitrocellulose membrane in parallel (dots V–VII). Membranes were washed subsequently and submitted to immunoblot analyses with anti-vimentin, anti-α-tubulin or anti-β-actin antibodies, respectively, due to the schematic shown. In contrast to α-tubulin and β-actin, only vimentin can be detected with its specific antibody on the nitrocellulose membrane, after overlaying ORF1p dots with the respective IF protein (dots I–III). This is indicating that only vimentin is binding to ORF1p.

    Article Snippet: Bovine α-tubulin and human β-actin were purchased from Cytoskeleton, Inc. (Denver, CO, USA).

    Techniques: In Situ, Recombinant, Immunofluorescence, Generated, Dot Blot, Overlay Assay, Blocking Assay, Incubation, Binding Assay

    ( a ) Effect of EMP2 on FAK phosphorylation. Levels of total FAK, p-FAK (Y397, Y407, Y861, and Y925), and β -actin were measured by Western blot analysis in the cell lines ARPE-19 and ARPE-19/EMP2 and were quantified by densitometry. ( b ) p-Src (Tyr

    Journal: Investigative ophthalmology & visual science

    Article Title: Functional Consequences of Interactions between FAK and Epithelial Membrane Protein 2 (EMP2)

    doi: 10.1167/iovs.08-3315

    Figure Lengend Snippet: ( a ) Effect of EMP2 on FAK phosphorylation. Levels of total FAK, p-FAK (Y397, Y407, Y861, and Y925), and β -actin were measured by Western blot analysis in the cell lines ARPE-19 and ARPE-19/EMP2 and were quantified by densitometry. ( b ) p-Src (Tyr

    Article Snippet: A mouse antibody recognizing human β -actin (clone 2A2.1) was from US Biological (Swampscott, MA).

    Techniques: Western Blot

    Effects of bile acids on the expression of HCV NS5B in GS4.1 cells. Unconjugated bile acid CDCA or DCA was incubated in 1- or 2-day-old semiconfluent GS4.1 cells for 48 h. Cell lysates were prepared after the incubation, and Western blot analysis was performed for the expression of HCV NS5B. As a loading control, Western blot analysis of β-actin was performed with the same samples.

    Journal: Journal of Virology

    Article Title: Bile Acids Promote the Expression of Hepatitis C Virus in Replicon-Harboring Cells ▿

    doi: 10.1128/JVI.00795-07

    Figure Lengend Snippet: Effects of bile acids on the expression of HCV NS5B in GS4.1 cells. Unconjugated bile acid CDCA or DCA was incubated in 1- or 2-day-old semiconfluent GS4.1 cells for 48 h. Cell lysates were prepared after the incubation, and Western blot analysis was performed for the expression of HCV NS5B. As a loading control, Western blot analysis of β-actin was performed with the same samples.

    Article Snippet: The monoclonal antibody specific for HCV NS5B or human β-actin was obtained from ViroStat (Portland, ME) or Cell Signaling Technology (Davers, MA), respectively.

    Techniques: Expressing, Incubation, Western Blot

    TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and β-actin.

    Journal: Molecular Cancer

    Article Title: Identification of the B-Raf/Mek/Erk MAP kinase pathway as a target for all-trans retinoic acid during skin cancer promotion

    doi: 10.1186/1476-4598-8-27

    Figure Lengend Snippet: TPA does not induce p38 and JNK phosphorylation . A . Paraffin sections of SENCAR mouse skin treated for 7 weeks with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with phospho-p38 and phospho-JNK antibodies followed by an Alexa-546-labeled secondary antibody and DAPI counterstain. All panels in A were photographed at 600× magnification. B . Western blots were run as in Fig. 4E for 7 week epidermal lysates and probed with antibodies for total p38, phospho-p38 (p-p38), total JNK, phospho-JNK (p-JNK) and β-actin.

    Article Snippet: Blots were blocked with 5% BSA for 1 h at room temperature, followed by incubation overnight with antibodies against phosphorylated forms of B-Raf (Ser 445), Mek1/2 (Ser 217/221), Erk1/2 (Thr202/Tyr204), SAPK/JNK (Thr183/Tyr185), and p38 MAPK (Thr180/Tyr182); and total B-Raf, c-Raf, Mek1/2, Erk1/2, p39 and SAPK/JNK; and human β-actin (all from Cell Signaling Technology, except for B-Raf and β-actin from Santa Cruz Biotechnology).

    Techniques: Labeling, Western Blot

    ATRA suppresses TPA induced phosphorylation of B-Raf, Mek1/2, and Erk1/2 . A . IHC staining of 3 week treated skin. Paraffin sections of mouse skin treated with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with pB-Raf, pMek1/2, and pERK1/2 antibodies followed by an Alexa-546-labeled secondary antibody. Sections were counterstaining with DAPI. All panels were photographed at 600× magnification. B . Epidermal lysates were prepared in RIPA buffer from SENCAR mouse skin treated with Acetone (Con), TPA, TPA+ATRA (T+A), or ATRA alone for 10 weeks. Total protein (15 μg per well) from pooled samples (n = 5) was run on 10% SDS-PAGE and probed with antibodies for total B-Raf, pB-Raf, total Mek1/2, pMek1/2, total Erk1/2, pERK1/2, and β-actin. C . Epidermal thickness during the time course. Thickness was measured from digital micrographs of H E stained skin sections as described in Methods. The symbols and error bars indicate the mean of 10 measurements ± SEM.

    Journal: Molecular Cancer

    Article Title: Identification of the B-Raf/Mek/Erk MAP kinase pathway as a target for all-trans retinoic acid during skin cancer promotion

    doi: 10.1186/1476-4598-8-27

    Figure Lengend Snippet: ATRA suppresses TPA induced phosphorylation of B-Raf, Mek1/2, and Erk1/2 . A . IHC staining of 3 week treated skin. Paraffin sections of mouse skin treated with Acetone (Control), TPA, TPA+ATRA, and ATRA alone were probed with pB-Raf, pMek1/2, and pERK1/2 antibodies followed by an Alexa-546-labeled secondary antibody. Sections were counterstaining with DAPI. All panels were photographed at 600× magnification. B . Epidermal lysates were prepared in RIPA buffer from SENCAR mouse skin treated with Acetone (Con), TPA, TPA+ATRA (T+A), or ATRA alone for 10 weeks. Total protein (15 μg per well) from pooled samples (n = 5) was run on 10% SDS-PAGE and probed with antibodies for total B-Raf, pB-Raf, total Mek1/2, pMek1/2, total Erk1/2, pERK1/2, and β-actin. C . Epidermal thickness during the time course. Thickness was measured from digital micrographs of H E stained skin sections as described in Methods. The symbols and error bars indicate the mean of 10 measurements ± SEM.

    Article Snippet: Blots were blocked with 5% BSA for 1 h at room temperature, followed by incubation overnight with antibodies against phosphorylated forms of B-Raf (Ser 445), Mek1/2 (Ser 217/221), Erk1/2 (Thr202/Tyr204), SAPK/JNK (Thr183/Tyr185), and p38 MAPK (Thr180/Tyr182); and total B-Raf, c-Raf, Mek1/2, Erk1/2, p39 and SAPK/JNK; and human β-actin (all from Cell Signaling Technology, except for B-Raf and β-actin from Santa Cruz Biotechnology).

    Techniques: Immunohistochemistry, Staining, Labeling, SDS Page

    SMAD3 Mediates Both the BTIC-Promoting and BTIC-Suppressing Programs of TGF-β (A) MDA-MB-231 and HCC-1954 first generation mammosphere cultures with and without addition of TGF-β. TGF-β was added to the media at the moment of cell seeding and mammospheres were allowed to form for 7 days. Note that this is not a quantitative assay. (B) Western blot showing SMAD2 phosphorylation levels upon TGF-β pathway induction. 7-day-old mammospheres were treated with exogenous TGF-β ligand for 1 hr. Total SMAD2/3 and β-actin levels were used as loading controls. (C) Comparison of TGF-β-dependent genes in MDA-MB-231 and HCC-1954 BTICs. MDA-MB-231 and HCC-1954 cells were grown as mammospheres for 7 days and treated with TGF-β for 1, 3, 6, and 24 hr, and gene expression profiling was performed using Illumina HumanHT-12 BeadChips. The plot shows the number of TGF-β-dependent genes in each BTIC at each time point (false discovery rate [FDR]

    Journal: Cell Reports

    Article Title: Context-Specific Effects of TGF-β/SMAD3 in Cancer Are Modulated by the Epigenome

    doi: 10.1016/j.celrep.2015.11.040

    Figure Lengend Snippet: SMAD3 Mediates Both the BTIC-Promoting and BTIC-Suppressing Programs of TGF-β (A) MDA-MB-231 and HCC-1954 first generation mammosphere cultures with and without addition of TGF-β. TGF-β was added to the media at the moment of cell seeding and mammospheres were allowed to form for 7 days. Note that this is not a quantitative assay. (B) Western blot showing SMAD2 phosphorylation levels upon TGF-β pathway induction. 7-day-old mammospheres were treated with exogenous TGF-β ligand for 1 hr. Total SMAD2/3 and β-actin levels were used as loading controls. (C) Comparison of TGF-β-dependent genes in MDA-MB-231 and HCC-1954 BTICs. MDA-MB-231 and HCC-1954 cells were grown as mammospheres for 7 days and treated with TGF-β for 1, 3, 6, and 24 hr, and gene expression profiling was performed using Illumina HumanHT-12 BeadChips. The plot shows the number of TGF-β-dependent genes in each BTIC at each time point (false discovery rate [FDR]

    Article Snippet: Membranes were blocked in 5% dried milk powder, 0.1% Tween-20 in PBS, and the following antibodies were used for protein detection: rabbit monoclonal against human phospho-SMAD2 (Ser 465/467) (Cell Signaling, 3108), rabbit polyclonal against human SMAD2/3 (Santa Cruz Biotechnology, sc-8332) and rabbit polyclonal against human β-actin (Abcam, ab8227).

    Techniques: Multiple Displacement Amplification, Western Blot, Expressing

    Expression of Elm1 and Cyr61 after serum stimulation of BALB/c 3T3 cells. 2 μg of poly(A) + RNA resolved electrophoretically were hybridized with the Elm1 ( A ) or Cyr61 ( B ) cDNA probe. The membrane was rehybridized with a human β-actin probe ( C ). poly(A) + RNA from quiescent cells (lane 1 ), cells stimulated with serum for 30 min (lane 2 ), and 3 h (lane 3 ) were loaded.

    Journal: The Journal of Experimental Medicine

    Article Title: Expression of the Elm1 Gene, a Novel Gene of the CCN (Connective Tissue Growth Factor, Cyr61/Cef10, and Neuroblastoma Overexpressed Gene) Family, Suppresses In Vivo Tumor Growth and Metastasis of K-1735 Murine Melanoma Cells

    doi:

    Figure Lengend Snippet: Expression of Elm1 and Cyr61 after serum stimulation of BALB/c 3T3 cells. 2 μg of poly(A) + RNA resolved electrophoretically were hybridized with the Elm1 ( A ) or Cyr61 ( B ) cDNA probe. The membrane was rehybridized with a human β-actin probe ( C ). poly(A) + RNA from quiescent cells (lane 1 ), cells stimulated with serum for 30 min (lane 2 ), and 3 h (lane 3 ) were loaded.

    Article Snippet: To confirm the amounts of mRNA loaded in each lane, the blots were hybridized afterwards with a human β-actin probe ( Clontech ).

    Techniques: Expressing

    ( A ) Southern blot analysis of DNA from various species using the Elm1 cDNA probe. Lane 1 , human; lane 2 , monkey; lane 3 , rat; lane 4 , mouse; lane 5 , dog; lane 6 , cow; lane 7 , rabbit; lane 8 , chicken; lane 9 , yeast. ( B ) Northern blot analysis of the Elm1 gene in various mouse tissues. Mouse multiple tissue Northern blot ( Clontech ) was hybridized with the Elm1 cDNA probe ( top ) or a human β-actin probe ( bottom ). Lane 1 , heart; lane 2 , brain; lane 3 , spleen; lane 4 , lung; lane 5 , liver; lane 6 , skeletal muscle; lane 7 , kidney; lane 8 , testis.

    Journal: The Journal of Experimental Medicine

    Article Title: Expression of the Elm1 Gene, a Novel Gene of the CCN (Connective Tissue Growth Factor, Cyr61/Cef10, and Neuroblastoma Overexpressed Gene) Family, Suppresses In Vivo Tumor Growth and Metastasis of K-1735 Murine Melanoma Cells

    doi:

    Figure Lengend Snippet: ( A ) Southern blot analysis of DNA from various species using the Elm1 cDNA probe. Lane 1 , human; lane 2 , monkey; lane 3 , rat; lane 4 , mouse; lane 5 , dog; lane 6 , cow; lane 7 , rabbit; lane 8 , chicken; lane 9 , yeast. ( B ) Northern blot analysis of the Elm1 gene in various mouse tissues. Mouse multiple tissue Northern blot ( Clontech ) was hybridized with the Elm1 cDNA probe ( top ) or a human β-actin probe ( bottom ). Lane 1 , heart; lane 2 , brain; lane 3 , spleen; lane 4 , lung; lane 5 , liver; lane 6 , skeletal muscle; lane 7 , kidney; lane 8 , testis.

    Article Snippet: To confirm the amounts of mRNA loaded in each lane, the blots were hybridized afterwards with a human β-actin probe ( Clontech ).

    Techniques: Southern Blot, Northern Blot

    Northern blot analysis of Elm1 transfectants using the Elm1 ( A ), Neo ( B ), and β-actin ( C ) probes. The size of exogenous and endogenous Elm1 transcripts are 1.5 and 5.0 kb, respectively. Lane 1 , K-1735 C-23; lane 2 , K-1735 M-2; lane 3 , M-2.Elm1. 23-1; lane 4 , M-2.Elm1.20-3; lane 5 , M-2.Elm1.0-1; lane 6 , M-2. Elm1.6-2; lane 7 , M-2. Elm1. 20-2; lane 8 , M-2.Elm1.8-3.

    Journal: The Journal of Experimental Medicine

    Article Title: Expression of the Elm1 Gene, a Novel Gene of the CCN (Connective Tissue Growth Factor, Cyr61/Cef10, and Neuroblastoma Overexpressed Gene) Family, Suppresses In Vivo Tumor Growth and Metastasis of K-1735 Murine Melanoma Cells

    doi:

    Figure Lengend Snippet: Northern blot analysis of Elm1 transfectants using the Elm1 ( A ), Neo ( B ), and β-actin ( C ) probes. The size of exogenous and endogenous Elm1 transcripts are 1.5 and 5.0 kb, respectively. Lane 1 , K-1735 C-23; lane 2 , K-1735 M-2; lane 3 , M-2.Elm1. 23-1; lane 4 , M-2.Elm1.20-3; lane 5 , M-2.Elm1.0-1; lane 6 , M-2. Elm1.6-2; lane 7 , M-2. Elm1. 20-2; lane 8 , M-2.Elm1.8-3.

    Article Snippet: To confirm the amounts of mRNA loaded in each lane, the blots were hybridized afterwards with a human β-actin probe ( Clontech ).

    Techniques: Northern Blot

    Csk expression and the kinase activity of synoviocytes. ( a ) Western blot analysis shows that Csk virus induced Csk expression in human synovial cells in an moi-dependent manner without significant changes in expression of c-Src protein. Fold increase in Csk expression normalized by β-actin is shown below each blot. The results shown are from a single experiment typical of 3 experiments yielding identical results. ( b ) Immunofluorescence staining using anti-Csk antibody shows that WT virus–infected synoviocytes expressed a very small amount of Csk protein. ( c ) Diffuse cytoplasmic staining in Csk virus–infected (moi = 100) synoviocytes. ( d ) In vitro kinase assay, using enolase as a substrate demonstrated that kinase activity of c-Src, Fyn, and c-Yes was remarkably repressed in Csk-overexpressing synoviocytes, but the expression of each protein was unaffected by Csk expression. Relative kinase activity of each kinase is shown below. The results were derived from 3 separate experiments. Suppression on each kinase was statistically significant (* P

    Journal: Journal of Clinical Investigation

    Article Title: Suppression of arthritic bone destruction by adenovirus-mediated csk gene transfer to synoviocytes and osteoclasts

    doi:

    Figure Lengend Snippet: Csk expression and the kinase activity of synoviocytes. ( a ) Western blot analysis shows that Csk virus induced Csk expression in human synovial cells in an moi-dependent manner without significant changes in expression of c-Src protein. Fold increase in Csk expression normalized by β-actin is shown below each blot. The results shown are from a single experiment typical of 3 experiments yielding identical results. ( b ) Immunofluorescence staining using anti-Csk antibody shows that WT virus–infected synoviocytes expressed a very small amount of Csk protein. ( c ) Diffuse cytoplasmic staining in Csk virus–infected (moi = 100) synoviocytes. ( d ) In vitro kinase assay, using enolase as a substrate demonstrated that kinase activity of c-Src, Fyn, and c-Yes was remarkably repressed in Csk-overexpressing synoviocytes, but the expression of each protein was unaffected by Csk expression. Relative kinase activity of each kinase is shown below. The results were derived from 3 separate experiments. Suppression on each kinase was statistically significant (* P

    Article Snippet: Human IL-6 probe was the PCR product of synoviocytes using the primers described previously , and human β-actin probe was obtained from Wako Pure Chemicals Industries.

    Techniques: Expressing, Activity Assay, Western Blot, Immunofluorescence, Staining, Infection, In Vitro, Kinase Assay, Derivative Assay

    In vivo expression of transgenes by adenovirus vectors. ( a ) In situ β-gal staining of the ankle synovial membrane, dissected 7 days after injection of LacZ virus, revealed a diffuse β-gal expression in the synoviocytes of rat ankles with adjuvant arthritis. ( b ) Multinucleated giant cells (arrowheads) at the bone/synovium interface were preferentially stained for β-gal. ( c ) These multinucleated cells (arrowheads) were positively stained for TRAP, a specific marker for osteoclasts. ( d ) In vivo expression of Csk protein in rat ankles detected by Western blotting ( n = 2 each day). Fold increase in Csk expression normalized by β-actin is shown below each blot. The results were derived from 3 separate experiments, with duplicates performed in each experiment.

    Journal: Journal of Clinical Investigation

    Article Title: Suppression of arthritic bone destruction by adenovirus-mediated csk gene transfer to synoviocytes and osteoclasts

    doi:

    Figure Lengend Snippet: In vivo expression of transgenes by adenovirus vectors. ( a ) In situ β-gal staining of the ankle synovial membrane, dissected 7 days after injection of LacZ virus, revealed a diffuse β-gal expression in the synoviocytes of rat ankles with adjuvant arthritis. ( b ) Multinucleated giant cells (arrowheads) at the bone/synovium interface were preferentially stained for β-gal. ( c ) These multinucleated cells (arrowheads) were positively stained for TRAP, a specific marker for osteoclasts. ( d ) In vivo expression of Csk protein in rat ankles detected by Western blotting ( n = 2 each day). Fold increase in Csk expression normalized by β-actin is shown below each blot. The results were derived from 3 separate experiments, with duplicates performed in each experiment.

    Article Snippet: Human IL-6 probe was the PCR product of synoviocytes using the primers described previously , and human β-actin probe was obtained from Wako Pure Chemicals Industries.

    Techniques: In Vivo, Expressing, In Situ, Staining, Injection, Marker, Western Blot, Derivative Assay

    Representative semiquantitative RT-PCR gel image demonstrating expression of testicular genes in calorie restricted ( CR ; n = 5) or control ( CON ; n = 5) young adult (∼12 years old) rhesus macaques. The housekeeping gene β-actin was used as a positive control and for normalizing images for analysis. No significant differences (mean ± SEM; P > 0.05) in gene expression were observed between the treatment groups

    Journal: Age

    Article Title: Impact of moderate calorie restriction on testicular morphology and endocrine function in adult rhesus macaques (Macaca mulatta)

    doi: 10.1007/s11357-013-9563-6

    Figure Lengend Snippet: Representative semiquantitative RT-PCR gel image demonstrating expression of testicular genes in calorie restricted ( CR ; n = 5) or control ( CON ; n = 5) young adult (∼12 years old) rhesus macaques. The housekeeping gene β-actin was used as a positive control and for normalizing images for analysis. No significant differences (mean ± SEM; P > 0.05) in gene expression were observed between the treatment groups

    Article Snippet: The PCR mixtures contained 5 μl of Taqman® Universal PCR Master Mix (Applied Biosystems), 0.3 μl of primer (300 nM final concentration; Invitrogen), 0.05 μl of human β-actin primers (50 nM final concentration; Applied Biosystems), 0.25 μl of probe (250 nM final concentration; IDT, Coralville, IA), and 2 μl cDNA.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Positive Control

    Representative semiquantitative RT-PCR gel image demonstrating expression of testicular genes in calorie restricted ( CR ; n = 3) or control ( CON ; n = 3) old adult (24–30 years old) rhesus macaques. The housekeeping gene β-actin was used as a positive control and for normalizing images for analysis. No significant differences (mean ± SEM; P > 0.05) in gene expression were observed between the treatment groups

    Journal: Age

    Article Title: Impact of moderate calorie restriction on testicular morphology and endocrine function in adult rhesus macaques (Macaca mulatta)

    doi: 10.1007/s11357-013-9563-6

    Figure Lengend Snippet: Representative semiquantitative RT-PCR gel image demonstrating expression of testicular genes in calorie restricted ( CR ; n = 3) or control ( CON ; n = 3) old adult (24–30 years old) rhesus macaques. The housekeeping gene β-actin was used as a positive control and for normalizing images for analysis. No significant differences (mean ± SEM; P > 0.05) in gene expression were observed between the treatment groups

    Article Snippet: The PCR mixtures contained 5 μl of Taqman® Universal PCR Master Mix (Applied Biosystems), 0.3 μl of primer (300 nM final concentration; Invitrogen), 0.05 μl of human β-actin primers (50 nM final concentration; Applied Biosystems), 0.25 μl of probe (250 nM final concentration; IDT, Coralville, IA), and 2 μl cDNA.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Positive Control

    Effect of chloroform fraction from Carpinus tschonoskii on the STAT1 signal in IFN-γ-stimulated HaCaT human keratinocytes. (A) Cells were pretreated with the indicated concentration of chloroform fraction of C. tschonoskii for 2 hr, and then the phosphorylation of STAT1 was determined in cells stimulated by IFN-γ (10 ng/ml) for 30 min. The levels of STAT1 and β-actin were identified using Western blotting analysis. (B) Cells were pretreated with the chloroform fraction (50 μg/ml) of C. tschonoskii for 2 hr, and then the translocation of STAT1 protein was determined in cells stimulated by IFN-γ (10 ng/ml) for 60 min. Immunofluorescence stain of STAT1 was stained with DyLight488-conjugated 2 nd antibody and the fluorescence was identified using confocal microscopy (FV500, OLYMPUS) and the images were acquired at constant conditions (PMT, gain, offset, magnification (20 × objective with zoom factor of 2.5) and resolution, etc.). These data are representative of three independent experiments.

    Journal: Toxicological Research

    Article Title: The Chloroform Fraction of Carpinus tschonoskii Leaves Inhibits the Production of Inflammatory Mediators in HaCaT Keratinocytes and RAW264.7 Macrophages

    doi: 10.5487/TR.2012.28.4.255

    Figure Lengend Snippet: Effect of chloroform fraction from Carpinus tschonoskii on the STAT1 signal in IFN-γ-stimulated HaCaT human keratinocytes. (A) Cells were pretreated with the indicated concentration of chloroform fraction of C. tschonoskii for 2 hr, and then the phosphorylation of STAT1 was determined in cells stimulated by IFN-γ (10 ng/ml) for 30 min. The levels of STAT1 and β-actin were identified using Western blotting analysis. (B) Cells were pretreated with the chloroform fraction (50 μg/ml) of C. tschonoskii for 2 hr, and then the translocation of STAT1 protein was determined in cells stimulated by IFN-γ (10 ng/ml) for 60 min. Immunofluorescence stain of STAT1 was stained with DyLight488-conjugated 2 nd antibody and the fluorescence was identified using confocal microscopy (FV500, OLYMPUS) and the images were acquired at constant conditions (PMT, gain, offset, magnification (20 × objective with zoom factor of 2.5) and resolution, etc.). These data are representative of three independent experiments.

    Article Snippet: Human β-actin primers were purchased from Bionex (Seoul, Korea).

    Techniques: Concentration Assay, Western Blot, Translocation Assay, Immunofluorescence, Staining, Fluorescence, Confocal Microscopy