Structured Review

Millipore sa β gal activity
Effect of ZER on intracellular ROS levels and SA- β -gal activity in UVA-irradiated HSF cells, (a) Cells were pretreated with ZER (0, 2, 4, or 8  μ M for 24 h) followed by irradiated with UVA in the absence or presence of 3 J/cm 2  UVA. The accumulation of UVA-induced ROS and SA- β -gal-positive cells was measured using fluorescence microscopy (200x magnification) as described. (b) The data was represented as fold change over control intracellular ROS levels. (c) The data was reported as fold increase over control SA- β -gal-positive cells. Results from three or more experiments were presented as mean ± SD, and the statistical significance was considered as ∗∗∗ p
Sa β Gal Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Zerumbone Exhibits Antiphotoaging and Dermatoprotective Properties in Ultraviolet A-Irradiated Human Skin Fibroblast Cells via the Activation of Nrf2/ARE Defensive Pathway"

Article Title: Zerumbone Exhibits Antiphotoaging and Dermatoprotective Properties in Ultraviolet A-Irradiated Human Skin Fibroblast Cells via the Activation of Nrf2/ARE Defensive Pathway

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2019/4098674

Effect of ZER on intracellular ROS levels and SA- β -gal activity in UVA-irradiated HSF cells, (a) Cells were pretreated with ZER (0, 2, 4, or 8  μ M for 24 h) followed by irradiated with UVA in the absence or presence of 3 J/cm 2  UVA. The accumulation of UVA-induced ROS and SA- β -gal-positive cells was measured using fluorescence microscopy (200x magnification) as described. (b) The data was represented as fold change over control intracellular ROS levels. (c) The data was reported as fold increase over control SA- β -gal-positive cells. Results from three or more experiments were presented as mean ± SD, and the statistical significance was considered as ∗∗∗ p
Figure Legend Snippet: Effect of ZER on intracellular ROS levels and SA- β -gal activity in UVA-irradiated HSF cells, (a) Cells were pretreated with ZER (0, 2, 4, or 8  μ M for 24 h) followed by irradiated with UVA in the absence or presence of 3 J/cm 2 UVA. The accumulation of UVA-induced ROS and SA- β -gal-positive cells was measured using fluorescence microscopy (200x magnification) as described. (b) The data was represented as fold change over control intracellular ROS levels. (c) The data was reported as fold increase over control SA- β -gal-positive cells. Results from three or more experiments were presented as mean ± SD, and the statistical significance was considered as ∗∗∗ p

Techniques Used: Activity Assay, Irradiation, Fluorescence, Microscopy

Related Articles

Fluorescence:

Article Title: Bmi1 limits dilated cardiomyopathy and heart failure by inhibiting cardiac senescence
Article Snippet: .. For fluorescence detection of SA-β-gal activity, cells (107 cell ml−1 ) were incubated with C12 FDG (fluorescein di-β-D -galactopyranoside; 33 μM; Sigma), a β-galactosidase substrate that generates a fluorescent product upon cleavage, for 60 min at 37 °C (ref. ). .. Cytochemical detection of senescent cells in vitro was determined in cells and fixed tissues with the Senescence β-Galactosidase Staining Kit (Cell Signaling).

Article Title: Cell size is a determinant of stem cell potential during aging
Article Snippet: To detect senescent HSCs, we used C12 FDG (#7188, Setareh Biotech) to measure the activity of the senescence biomarker SA-β-galactosidase in un-fixed HSCs by flow cytometry ( ). .. For fluorescence detection of SA-β-galactosidase activity, 107 BM cell/mL were incubated with 30 μM chloroquine (#C6628-25G, Sigma Aldrich) in IMDM supplemented with 2 % FBS at 37°C for 30 min. 32 μM C12 -FDG, a β-galactosidase substrate that generates a fluorescent product upon cleavage, was added for 30 min in a 37°C water bath. .. Cells were washed with 2 % IMDM at 4°C, lineage positive cells were depleted, lin-negative cells stained with HSC antibodies as described above and analyzed by flow cytometry.

Activity Assay:

Article Title: Bmi1 limits dilated cardiomyopathy and heart failure by inhibiting cardiac senescence
Article Snippet: .. For fluorescence detection of SA-β-gal activity, cells (107 cell ml−1 ) were incubated with C12 FDG (fluorescein di-β-D -galactopyranoside; 33 μM; Sigma), a β-galactosidase substrate that generates a fluorescent product upon cleavage, for 60 min at 37 °C (ref. ). .. Cytochemical detection of senescent cells in vitro was determined in cells and fixed tissues with the Senescence β-Galactosidase Staining Kit (Cell Signaling).

Article Title: Inflammation-induced Gro1 triggers senescence in neuronal progenitors: effects of estradiol
Article Snippet: The following primary antibodies were used: Tuj-1 (Stem Cell Technology, cat# 01409 or Abcam, cat# ab182-07), Ng2 (Millipore, cat# AB5320), GFAP (Millipore, cat# MAB3402 or Abcam, cat# ab7260), Ki67 (Abcam, cat#ab15580), DCX (Abcam cat#ab18723), and p16 (Santa Cruz cat# sc-1207 or sc-1661). .. SA-β-galactosidase activity SA-β-galactosidase enzymatic activity was assayed in vitro using a β-gal staining kit (Senescence Cell Staining Kit, Sigma-Aldrich) according to the manual. .. Briefly, 10,000 cells were plated in 12-well plates, treated for the indicated times, washed with PBS (pH 6.0), fixed, and stained with 5-bromo-4-chloro-3-indolyl-h-d -galactopyranoside (X-Gal) overnight at 37 °C.

Article Title: Effect of Advanced Glycation End Products on Oxidative Stress and Senescence of Trabecular Meshwork Cells
Article Snippet: Next, flow cytometric analysis was performed (Cytomics 500FC; Beckman Coulter, Miami, FL, USA) at fluorescence emission wavelengths less than 530 nm and greater than 575 nm. .. Assessment of cellular senescence A commercial senescence-associated β-galactosidase kit (SA-β-gal, Sigma) was used to measure β-galactosidase activity, which is characteristic of senescent cells [ , ]. .. SA-β-gal is used to assess abnormal enzymatic activity observed in aging.

Article Title: Cell size is a determinant of stem cell potential during aging
Article Snippet: To detect senescent HSCs, we used C12 FDG (#7188, Setareh Biotech) to measure the activity of the senescence biomarker SA-β-galactosidase in un-fixed HSCs by flow cytometry ( ). .. For fluorescence detection of SA-β-galactosidase activity, 107 BM cell/mL were incubated with 30 μM chloroquine (#C6628-25G, Sigma Aldrich) in IMDM supplemented with 2 % FBS at 37°C for 30 min. 32 μM C12 -FDG, a β-galactosidase substrate that generates a fluorescent product upon cleavage, was added for 30 min in a 37°C water bath. .. Cells were washed with 2 % IMDM at 4°C, lineage positive cells were depleted, lin-negative cells stained with HSC antibodies as described above and analyzed by flow cytometry.

Incubation:

Article Title: Bmi1 limits dilated cardiomyopathy and heart failure by inhibiting cardiac senescence
Article Snippet: .. For fluorescence detection of SA-β-gal activity, cells (107 cell ml−1 ) were incubated with C12 FDG (fluorescein di-β-D -galactopyranoside; 33 μM; Sigma), a β-galactosidase substrate that generates a fluorescent product upon cleavage, for 60 min at 37 °C (ref. ). .. Cytochemical detection of senescent cells in vitro was determined in cells and fixed tissues with the Senescence β-Galactosidase Staining Kit (Cell Signaling).

Article Title: Cell size is a determinant of stem cell potential during aging
Article Snippet: To detect senescent HSCs, we used C12 FDG (#7188, Setareh Biotech) to measure the activity of the senescence biomarker SA-β-galactosidase in un-fixed HSCs by flow cytometry ( ). .. For fluorescence detection of SA-β-galactosidase activity, 107 BM cell/mL were incubated with 30 μM chloroquine (#C6628-25G, Sigma Aldrich) in IMDM supplemented with 2 % FBS at 37°C for 30 min. 32 μM C12 -FDG, a β-galactosidase substrate that generates a fluorescent product upon cleavage, was added for 30 min in a 37°C water bath. .. Cells were washed with 2 % IMDM at 4°C, lineage positive cells were depleted, lin-negative cells stained with HSC antibodies as described above and analyzed by flow cytometry.

Staining:

Article Title: CTGF Attenuates Tendon-Derived Stem/Progenitor Cell Aging
Article Snippet: TSPCs were transfected for 72 h, and the transfection mixture was replaced with culture medium. .. 2.5. β -Galactosidase Staining To study the effect of CTGF on cell senescence, the β -galactosidase (β -gal) assay was performed using the SA-β -gal staining kit (Sigma, USA). .. TSPCs were grown in 12-well plates for 24 h; then, cells were cultured with or without 250 ng/mL CTGF.

Article Title: Inflammation-induced Gro1 triggers senescence in neuronal progenitors: effects of estradiol
Article Snippet: The following primary antibodies were used: Tuj-1 (Stem Cell Technology, cat# 01409 or Abcam, cat# ab182-07), Ng2 (Millipore, cat# AB5320), GFAP (Millipore, cat# MAB3402 or Abcam, cat# ab7260), Ki67 (Abcam, cat#ab15580), DCX (Abcam cat#ab18723), and p16 (Santa Cruz cat# sc-1207 or sc-1661). .. SA-β-galactosidase activity SA-β-galactosidase enzymatic activity was assayed in vitro using a β-gal staining kit (Senescence Cell Staining Kit, Sigma-Aldrich) according to the manual. .. Briefly, 10,000 cells were plated in 12-well plates, treated for the indicated times, washed with PBS (pH 6.0), fixed, and stained with 5-bromo-4-chloro-3-indolyl-h-d -galactopyranoside (X-Gal) overnight at 37 °C.

Article Title: Progerin Expression Induces Inflammation, Oxidative Stress and Senescence in Human Coronary Endothelial Cells
Article Snippet: Cellular Senescence AssayNuclear foci secondary to DNA Double-Strand Breaks (DSBs) were visualized by immunofluorescence with antibodies directed against Ser139-phosphorylated histone variant H2A (γ-H2AX, 05-636, Merk, Sigma-Aldrich) in transduced HCAECs. .. Nuclear DNA was stained with di-amidino-2-phenylindole hydrochloride staining (DAPI, Sigma-Aldrich). .. The blue staining produced by SA-β-galactosidase’s hydrolysis of 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-Gal, Sigma-Aldrich) was used as a biomarker of cellular senescence [ ].

β-Gal Assay:

Article Title: CTGF Attenuates Tendon-Derived Stem/Progenitor Cell Aging
Article Snippet: TSPCs were transfected for 72 h, and the transfection mixture was replaced with culture medium. .. 2.5. β -Galactosidase Staining To study the effect of CTGF on cell senescence, the β -galactosidase (β -gal) assay was performed using the SA-β -gal staining kit (Sigma, USA). .. TSPCs were grown in 12-well plates for 24 h; then, cells were cultured with or without 250 ng/mL CTGF.

In Vitro:

Article Title: Inflammation-induced Gro1 triggers senescence in neuronal progenitors: effects of estradiol
Article Snippet: The following primary antibodies were used: Tuj-1 (Stem Cell Technology, cat# 01409 or Abcam, cat# ab182-07), Ng2 (Millipore, cat# AB5320), GFAP (Millipore, cat# MAB3402 or Abcam, cat# ab7260), Ki67 (Abcam, cat#ab15580), DCX (Abcam cat#ab18723), and p16 (Santa Cruz cat# sc-1207 or sc-1661). .. SA-β-galactosidase activity SA-β-galactosidase enzymatic activity was assayed in vitro using a β-gal staining kit (Senescence Cell Staining Kit, Sigma-Aldrich) according to the manual. .. Briefly, 10,000 cells were plated in 12-well plates, treated for the indicated times, washed with PBS (pH 6.0), fixed, and stained with 5-bromo-4-chloro-3-indolyl-h-d -galactopyranoside (X-Gal) overnight at 37 °C.

Mouse Assay:

Article Title: RhoA/ROCK inhibition improves the beneficial effects of glucocorticoid treatment in dystrophic muscle: implications for stem cell depletion
Article Snippet: .. MDSCs from 6-week-old dKO mice were cultured in the presence of prednisolone (Sigma, St Louis, MO) (10 µg/ml) ( , ) for 7 days. .. Prednisolone treatment of the cells was refreshed every 2 days by changing the medium (PM). dKO MDSCs were then fixed with cold methanol and analysed for their expression of p-4E-BP1 and SA-β-gal.

Cell Culture:

Article Title: RhoA/ROCK inhibition improves the beneficial effects of glucocorticoid treatment in dystrophic muscle: implications for stem cell depletion
Article Snippet: .. MDSCs from 6-week-old dKO mice were cultured in the presence of prednisolone (Sigma, St Louis, MO) (10 µg/ml) ( , ) for 7 days. .. Prednisolone treatment of the cells was refreshed every 2 days by changing the medium (PM). dKO MDSCs were then fixed with cold methanol and analysed for their expression of p-4E-BP1 and SA-β-gal.

Expressing:

Article Title: TLR8 signaling enhances tumor immunity by preventing tumor-induced T-cell senescence
Article Snippet: TLR ligands included: Pam3CSK4 (200 ng/ml), Poly (I:C) (25 μg/ml), LPS (100 ng/ml), Flagellin (10 μg/ml), Loxoribine (500 μM), ssRNA40/LyoVec (3 μg/ml) (Invivogen, San Diego, CA), and oligonucleotides CpG-B (3 μg/ml), Poly-T3 (3 μg/ml) and Poly-G3 (3 μg/ml) (synthesized by Invitrogen, Carlsbad, CA); neutralizing antibodies included: anti-IL-10 (30 μg/ml) (Clone JES3-19F1, BD Biosciences) and antihuman LAP (TGF- β1 10 μg/ml), anti-TCRαβ 10 μg/ml), anti-PDL-1 (10 μg/ml) (R & D Systems, Minneapolis, MN), anti-MHC-class II (Hybridoma supernatants, 200 μg/ml); ATM inhibitor KU55933 (20 μM, Tocris Bioscience); cAMP and PKA inhibitors included: 2′,5′-Dideoxyadenosine (7ddA, 320 μM) and Dihydrochloride (H89, 20 μM) (Calbiochemistry, San Diego, CA). .. For forskolin and 3-isobutyl-1-methylxanthine (IBMX) treatment experiments, MCF7, M628, or PC3 tumor cells were pretreated with forskolin (50 μM, Calbiochemistry) or IBMX (200 μM, Sigma) for 2 days, then co-cultured with anti-CD3-activated CD4+ T cells and detected for SA-β-Gal expression as described above. .. In some experiments, anti-CD3-activated CD4+ T cells were directly cultured in the presence of forskolin (0, 0.5, 1, 2 μM) for 5 days and then detected for SA-β-Gal expression.

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  • 93
    Millipore β gal activity
    T-cell-specific stimulation of <t>β-Gal</t> expression mediated by the hCD2 LCR in cell pools. HT1080, Jurkat, and primary human T cells were transduced with pHIV/CK-3 or with pHIV/CK-4 at an MOI of 10. Transduced cells were selected with G418. Cell extracts were prepared by freeze-thaw cycles. β-Gal expression in the cell pools was quantified and corrected for the protein concentration of the samples. All assays were carried out at least three times with freshly prepared cell extracts of exponentially growing cells. Standard deviations are indicated.
    β Gal Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β gal activity/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    β gal activity - by Bioz Stars, 2021-07
    93/100 stars
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    94
    Millipore rabbit anti β gal
    Effects of chronic pain on Wnt activity in ventral hippocampus. (A) Double-immunostaining of <t>Nestin/β-gal</t> and Western-blotting of β-gal in the ventral hippocampus of sham-injured and SNI-treated Topgal mice at 21 dpi. (B) Western-blotting of β-catenin and Axin2 in the ventral hippocampus of sham-injured and SNI-treated mice at 21 dpi. (C) Double-immunostaining and quantification of Nestin/β-gal in the SVZ of sham-injured and SNI-treated Topgal mice at 21 dpi. Inserts in (A) are magnified typical Nestin/β-gal-positive cells in each group, which were pointed by arrows. Bars = 25 μm in (A) and 50 μm in (C) . Values represent mean ± SE. Unpaired, two tailed Student’s t -tests were performed in (B,C) . * P
    Rabbit Anti β Gal, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti β gal/product/Millipore
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti β gal - by Bioz Stars, 2021-07
    94/100 stars
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    94
    Millipore β galactosidase sa β gal activity
    miR-34a induces lung fibroblast senescence. ( A – D ) Human lung fibroblasts MRC-5 were transfected with 50 nM control mimics or mimics for miR-34a. Three days after transfection, senescence–associated <t>β-galactosidase</t> <t>(SA-β-gal)</t> activities in the cells were determined ( A and B ). Levels of p21, plasminogen activator inhibitor-1 (PAI-1), nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4), and hyaluronan synthase (HAS2) were determined by real-time PCR ( C and D ) ( n = 3; mean ± SD). * P
    β Galactosidase Sa β Gal Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β galactosidase sa β gal activity/product/Millipore
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    β galactosidase sa β gal activity - by Bioz Stars, 2021-07
    94/100 stars
      Buy from Supplier

    93
    Millipore β gal
    YbeY does not comigrate with ribosomes. Cultures of E. coli ) ybeY and rpsT were grown in LB at 37°C to an OD 600 of 0.4 and then cooled down on ice for 15 min to allow runoff of nascent peptides. Next, lysates were prepared and 13 OD 260 units was separated through a 5 to 25% sucrose gradient by ultracentrifugation. Gradients were fractionated from top to bottom, and OD 260 was determined. The presence of YbeY, S20, and <t>β-gal</t> in the different fractions was determined by TCA concentration, SDS-PAGE separation, and immunoblotting as described in Materials and Methods.
    β Gal, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β gal/product/Millipore
    Average 93 stars, based on 1 article reviews
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    β gal - by Bioz Stars, 2021-07
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    T-cell-specific stimulation of β-Gal expression mediated by the hCD2 LCR in cell pools. HT1080, Jurkat, and primary human T cells were transduced with pHIV/CK-3 or with pHIV/CK-4 at an MOI of 10. Transduced cells were selected with G418. Cell extracts were prepared by freeze-thaw cycles. β-Gal expression in the cell pools was quantified and corrected for the protein concentration of the samples. All assays were carried out at least three times with freshly prepared cell extracts of exponentially growing cells. Standard deviations are indicated.

    Journal: Journal of Virology

    Article Title: Locus Control Region of the Human CD2 Gene in a Lentivirus Vector Confers Position-Independent Transgene Expression

    doi: 10.1128/JVI.75.10.4641-4648.2001

    Figure Lengend Snippet: T-cell-specific stimulation of β-Gal expression mediated by the hCD2 LCR in cell pools. HT1080, Jurkat, and primary human T cells were transduced with pHIV/CK-3 or with pHIV/CK-4 at an MOI of 10. Transduced cells were selected with G418. Cell extracts were prepared by freeze-thaw cycles. β-Gal expression in the cell pools was quantified and corrected for the protein concentration of the samples. All assays were carried out at least three times with freshly prepared cell extracts of exponentially growing cells. Standard deviations are indicated.

    Article Snippet: Units of active β-Gal were determined from a standard curve of β-Gal activity versus protein concentration, using purified β-Gal (Sigma, St. Louis, Mo.).

    Techniques: Expressing, Transduction, Protein Concentration

    LCR-mediated β-Gal gene expression in individual primary T-cell clones. Primary human T cells were transduced with pHIV/CK-3 at an MOI of 10. Transduced cells were selected with G418. The β-Gal activity in individual clones was quantified and corrected for the protein concentration in the samples. The average expression level is indicated by a horizontal line. Each dot represents the average β-Gal activity from three independent assays.

    Journal: Journal of Virology

    Article Title: Locus Control Region of the Human CD2 Gene in a Lentivirus Vector Confers Position-Independent Transgene Expression

    doi: 10.1128/JVI.75.10.4641-4648.2001

    Figure Lengend Snippet: LCR-mediated β-Gal gene expression in individual primary T-cell clones. Primary human T cells were transduced with pHIV/CK-3 at an MOI of 10. Transduced cells were selected with G418. The β-Gal activity in individual clones was quantified and corrected for the protein concentration in the samples. The average expression level is indicated by a horizontal line. Each dot represents the average β-Gal activity from three independent assays.

    Article Snippet: Units of active β-Gal were determined from a standard curve of β-Gal activity versus protein concentration, using purified β-Gal (Sigma, St. Louis, Mo.).

    Techniques: Expressing, Clone Assay, Transduction, Activity Assay, Protein Concentration

    Effects of chronic pain on Wnt activity in ventral hippocampus. (A) Double-immunostaining of Nestin/β-gal and Western-blotting of β-gal in the ventral hippocampus of sham-injured and SNI-treated Topgal mice at 21 dpi. (B) Western-blotting of β-catenin and Axin2 in the ventral hippocampus of sham-injured and SNI-treated mice at 21 dpi. (C) Double-immunostaining and quantification of Nestin/β-gal in the SVZ of sham-injured and SNI-treated Topgal mice at 21 dpi. Inserts in (A) are magnified typical Nestin/β-gal-positive cells in each group, which were pointed by arrows. Bars = 25 μm in (A) and 50 μm in (C) . Values represent mean ± SE. Unpaired, two tailed Student’s t -tests were performed in (B,C) . * P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Anxiety Specific Response and Contribution of Active Hippocampal Neural Stem Cells to Chronic Pain Through Wnt/β-Catenin Signaling in Mice

    doi: 10.3389/fnmol.2018.00296

    Figure Lengend Snippet: Effects of chronic pain on Wnt activity in ventral hippocampus. (A) Double-immunostaining of Nestin/β-gal and Western-blotting of β-gal in the ventral hippocampus of sham-injured and SNI-treated Topgal mice at 21 dpi. (B) Western-blotting of β-catenin and Axin2 in the ventral hippocampus of sham-injured and SNI-treated mice at 21 dpi. (C) Double-immunostaining and quantification of Nestin/β-gal in the SVZ of sham-injured and SNI-treated Topgal mice at 21 dpi. Inserts in (A) are magnified typical Nestin/β-gal-positive cells in each group, which were pointed by arrows. Bars = 25 μm in (A) and 50 μm in (C) . Values represent mean ± SE. Unpaired, two tailed Student’s t -tests were performed in (B,C) . * P

    Article Snippet: For immunostaining, the sections were blocked by 0.01 M phosphate buffered saline (PBS) containing 0.3% Triton X-100 and 3% bovine serum albumin (BSA) for 1 h. Primary antibodies were used as following: guinea pig anti-DCX (1:500, Millipore), rat anti-BrdU (1:200, Abcam), rabbit anti-NeuN (1:500, TEMECULA), goat anti-Nestin (1:500, Santa Cruz), rabbit anti-β-gal (1:500, MP), anti-β-catenin (1:200, Millipore).

    Techniques: Activity Assay, Double Immunostaining, Western Blot, Mouse Assay, Two Tailed Test

    Effects of Fluoxetine treatment on the Wnt activity in hippocampus. (A) Double-immunostaining of Nestin/β-gal in sham mice treated with saline (Sham+S), sham mice treated with Fluoxetine (Sham+F), SNI mice treated with saline (SNI+S), and SNI mice treated with Fluoxetine (SNI+F) at 21 dpi. Notice the significant increase of radial Nestin-positive cells in Fluoxetine treated mice. (B) Western-blotting of β-gal and β-catenin in Topgal mice with the following treatments: sham injury plus saline (Sham+S), sham injury plus Fluoxetine (Sham+F), SNI plus saline (SNI+S) and SNI plus Fluoxetine (SNI+F). Notice that Fluoxetine treatment significantly increased the expression level of β-gal and β-catenin in both sham and SNI-treated mice, as compared to corresponding saline controls. S, saline. F, Fluoxetine. Inserts in (A) are typical double-stained cells in each group, which were pointed by arrows. Bars = 50 μm. Values represent mean ± SE. Unpaired, two tailed Student’s t -tests were performed in (A,B) . * P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Anxiety Specific Response and Contribution of Active Hippocampal Neural Stem Cells to Chronic Pain Through Wnt/β-Catenin Signaling in Mice

    doi: 10.3389/fnmol.2018.00296

    Figure Lengend Snippet: Effects of Fluoxetine treatment on the Wnt activity in hippocampus. (A) Double-immunostaining of Nestin/β-gal in sham mice treated with saline (Sham+S), sham mice treated with Fluoxetine (Sham+F), SNI mice treated with saline (SNI+S), and SNI mice treated with Fluoxetine (SNI+F) at 21 dpi. Notice the significant increase of radial Nestin-positive cells in Fluoxetine treated mice. (B) Western-blotting of β-gal and β-catenin in Topgal mice with the following treatments: sham injury plus saline (Sham+S), sham injury plus Fluoxetine (Sham+F), SNI plus saline (SNI+S) and SNI plus Fluoxetine (SNI+F). Notice that Fluoxetine treatment significantly increased the expression level of β-gal and β-catenin in both sham and SNI-treated mice, as compared to corresponding saline controls. S, saline. F, Fluoxetine. Inserts in (A) are typical double-stained cells in each group, which were pointed by arrows. Bars = 50 μm. Values represent mean ± SE. Unpaired, two tailed Student’s t -tests were performed in (A,B) . * P

    Article Snippet: For immunostaining, the sections were blocked by 0.01 M phosphate buffered saline (PBS) containing 0.3% Triton X-100 and 3% bovine serum albumin (BSA) for 1 h. Primary antibodies were used as following: guinea pig anti-DCX (1:500, Millipore), rat anti-BrdU (1:200, Abcam), rabbit anti-NeuN (1:500, TEMECULA), goat anti-Nestin (1:500, Santa Cruz), rabbit anti-β-gal (1:500, MP), anti-β-catenin (1:200, Millipore).

    Techniques: Activity Assay, Double Immunostaining, Mouse Assay, Western Blot, Expressing, Staining, Two Tailed Test

    miR-34a induces lung fibroblast senescence. ( A – D ) Human lung fibroblasts MRC-5 were transfected with 50 nM control mimics or mimics for miR-34a. Three days after transfection, senescence–associated β-galactosidase (SA-β-gal) activities in the cells were determined ( A and B ). Levels of p21, plasminogen activator inhibitor-1 (PAI-1), nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4), and hyaluronan synthase (HAS2) were determined by real-time PCR ( C and D ) ( n = 3; mean ± SD). * P

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: miR-34a Inhibits Lung Fibrosis by Inducing Lung Fibroblast Senescence

    doi: 10.1165/rcmb.2016-0163OC

    Figure Lengend Snippet: miR-34a induces lung fibroblast senescence. ( A – D ) Human lung fibroblasts MRC-5 were transfected with 50 nM control mimics or mimics for miR-34a. Three days after transfection, senescence–associated β-galactosidase (SA-β-gal) activities in the cells were determined ( A and B ). Levels of p21, plasminogen activator inhibitor-1 (PAI-1), nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4), and hyaluronan synthase (HAS2) were determined by real-time PCR ( C and D ) ( n = 3; mean ± SD). * P

    Article Snippet: Cellular senescence was evaluated by determining cellular senescence–associated β-galactosidase (SA-β-gal) activity with either the Cellular Senescence Assay Kit (Millipore, Temecula, CA) or the MarkerGene Cellular Senescence Microtiterplate Assay Kit (Marker Gene Technologies, Eugene, OR) according to the manufacturer’s instructions.

    Techniques: Transfection, Real-time Polymerase Chain Reaction

    miR-34a knockdown inhibits senescence of lung fibroblasts. ( A and B ) Human lung fibroblasts MRC-5 were treated with low-dose bleomycin (0.01 U/ml) for 2 days. ( A ) SA-β-gal activities in the cells were measured, and ( B ) levels of miR-34a were determined by real-time PCR ( n = 3; mean ± SD). ** P

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: miR-34a Inhibits Lung Fibrosis by Inducing Lung Fibroblast Senescence

    doi: 10.1165/rcmb.2016-0163OC

    Figure Lengend Snippet: miR-34a knockdown inhibits senescence of lung fibroblasts. ( A and B ) Human lung fibroblasts MRC-5 were treated with low-dose bleomycin (0.01 U/ml) for 2 days. ( A ) SA-β-gal activities in the cells were measured, and ( B ) levels of miR-34a were determined by real-time PCR ( n = 3; mean ± SD). ** P

    Article Snippet: Cellular senescence was evaluated by determining cellular senescence–associated β-galactosidase (SA-β-gal) activity with either the Cellular Senescence Assay Kit (Millipore, Temecula, CA) or the MarkerGene Cellular Senescence Microtiterplate Assay Kit (Marker Gene Technologies, Eugene, OR) according to the manufacturer’s instructions.

    Techniques: Real-time Polymerase Chain Reaction

    YbeY does not comigrate with ribosomes. Cultures of E. coli ) ybeY and rpsT were grown in LB at 37°C to an OD 600 of 0.4 and then cooled down on ice for 15 min to allow runoff of nascent peptides. Next, lysates were prepared and 13 OD 260 units was separated through a 5 to 25% sucrose gradient by ultracentrifugation. Gradients were fractionated from top to bottom, and OD 260 was determined. The presence of YbeY, S20, and β-gal in the different fractions was determined by TCA concentration, SDS-PAGE separation, and immunoblotting as described in Materials and Methods.

    Journal: Journal of Bacteriology

    Article Title: The Heat Shock Protein YbeY Is Required for Optimal Activity of the 30S Ribosomal Subunit ▿

    doi: 10.1128/JB.00448-10

    Figure Lengend Snippet: YbeY does not comigrate with ribosomes. Cultures of E. coli ) ybeY and rpsT were grown in LB at 37°C to an OD 600 of 0.4 and then cooled down on ice for 15 min to allow runoff of nascent peptides. Next, lysates were prepared and 13 OD 260 units was separated through a 5 to 25% sucrose gradient by ultracentrifugation. Gradients were fractionated from top to bottom, and OD 260 was determined. The presence of YbeY, S20, and β-gal in the different fractions was determined by TCA concentration, SDS-PAGE separation, and immunoblotting as described in Materials and Methods.

    Article Snippet: Mouse monoclonal antibodies specific for luciferase, β-gal, and FLAG×3 (Sigma) were used as primary antibodies, and horseradish peroxidase-conjugated anti-mouse IgG (Sigma) was used as the secondary antibody.

    Techniques: Concentration Assay, SDS Page