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Cell Signaling Technology Inc anti rabbit
Anti Rabbit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 727 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti rabbit/product/Cell Signaling Technology Inc
Average 92 stars, based on 727 article reviews
Price from $9.99 to $1999.99
anti rabbit - by Bioz Stars, 2020-09
92/100 stars

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Western Blot:

Article Title: SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α
Article Snippet: .. Primary antibodies used were polyclonal anti-PARP-1 (89 and 116 KDa), anti-rabbits, 1 : 1000 in WB (Cell Signalling, Danvers, MA, USA), polyclonal anti-SGK-1 (50 kDa) anti-rabbits, 1 : 1000 in WB (UpState in Millipore), polyclonal anti-AKT-1 , (60 kDa) anti-rabbits, 1 : 1000 in WB (Cell Signalling), polyclonal p-AK T-1 (Ser 473) (60 kDa), anti-rabbits, 1 : 1000 in WB (Cell Signalling), monoclonal anti-caspase- 3 (clone 4-1-1-18) (32 and 17 KDa) anti-mouse 1 : 1000 in WB (UpState in Millipore), monoclonal anti-tubulin (clone DM 1 A) (55 kDa) anti-mouse 1 : 10 000 in WB (Sigma, Milan, Italy), anti-p-NDRG1 (Thr346 and Ser 330) (46–48 kDa) anti-rabbits, 1 : 1000 in WB (Cell Signalling). .. Cell treatments Insulin (Sigma): 10−7 M for 30 min. C 2 -Ceramide/C 6 -ceramide/C 2 -Dihydroceramide (BIOMOL in Enzo Life Science, Farmingdale, NY, USA): dissolved in DMSO (vehicle), then diluted into serum-free DMEM at the indicated concentrations and briefly sonicated, concentration used in the experiment is 50 μ M, in serum-free medium in 0.1% BSA RIA Grade (Sigma) for 48 h. TNF-α (Sigma): 100 ng/ml for 72 h. FumonisinB1 (Cayman Chemical Company, Ann Arbor, MA, USA): 100 μ M, 30 min before the stimulation with TNF-α .

other:

Article Title: Cox-2 Inhibition Protects against Hypoxia/Reoxygenation-Induced Cardiomyocyte Apoptosis via Akt-Dependent Enhancement of iNOS Expression
Article Snippet: Primary antibodies against Bcl2-like protein 4 (Bax, 1 : 1000, Cat. number 2772S), Bcl2 (1 : 1000, Cat. number 2870S), glyceraldehyde 3-phosphate dehydrogenase (GADPH, 1 : 1000, Cat. number 5174S), inhibitor of kappa B α (Iκ Bα , 1 : 1000, Cat. number 4814S), phospho-Iκ Bα (1 : 1000, Cat. number 9246S), p65 (1 : 1000, Cat. number 4764S), phospho-p65 (1 : 1000, Cat. number 3033S), total caspase-3 (1 : 1000, Cat. number 9665S), cleaved caspase-3 (1 : 1000, Cat. number 9661S), total Akt (1 : 1000, Cat. number 9272S), phosphor-Akt (1 : 1000, T450 Cat. number 9267S), and horseradish peroxidase-conjugated anti-mouse (Cat. number 7076S) or anti-rabbit (Cat. number 7074S) secondary antibodies (1 : 3000) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Article Title: Omega-3 Fatty Acids-Enriched Fish Oil Activates AMPK/PGC-1α Signaling and Prevents Obesity-Related Skeletal Muscle Wasting
Article Snippet: Anti-rabbit (7074S; 1:5000 dilution) (Cell Signaling Technology) and anti-mouse (7076S; 1:5000 dilution) (Cell Signaling Technology) horseradish peroxidase-conjugated secondary antibodies (anti)were then used to probe the membranes.

Article Title: Estrogen receptor (ER) was regulated by RNPC1 stabilizing mRNA in ER positive breast cancer
Article Snippet: The anti-rabbit and anti-mouse secondary antibodies were from Cell Signaling technology.

Article Title: H3K27 Demethylation at the Proviral Promoter Sensitizes Latent HIV to the Effects of Vorinostat in Ex Vivo Cultures of Resting CD4+ T Cells
Article Snippet: The following antibodies were used: anti-histone H3, anti-H3K27me3 (trimethylated histone 3 at lysine 27), anti-RNA polymerase II (Pol II), and anti-hemagglutinin (HA) (Abcam); anti-EZH2, anti-H3K27me2, anti-EED (embryonic ectoderm development gene), anti-RING1B, anti-BMI1, anti-H3K27ac, and anti-phospho-Ser2 Pol II (Active Motif); anti-H3K9me3 and anti-H3Ac (Millipore); and anti-HA, anti-rabbit, and anti-mouse immunoglobulin G (Cell Signaling).

Article Title: Effects of Chronic Overload on Muscle Hypertrophy and mTOR Signaling in Young Adult and Aged Rats
Article Snippet: Anti-rabbit and anti-mouse horseradish peroxidase–conjugated secondary antibodies were purchased from Cell Signaling Technologies.

Article Title: Hsp70 Cochaperones HspBP1 and BAG-1M Differentially Regulate Steroid Hormone Receptor Function
Article Snippet: Further antibodies were anti-CHIP (Calbiochem, San Diego, USA), anti-HA-HRP (3F10; Roche Diagnostics, Mannheim, Germany), anti-p23 (MA3-414; Affinity Bio Reagents, Golden, USA), anti-His (ab14923-100; Abcam, Massachusetts, USA), anti-HspBP1 (MAB-10201, Biozol, Eching, Germany), and anti-rabbit (Cell Signaling, Massachusetts, USA).

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  • 88
    Cell Signaling Technology Inc rabbit anti p fak
    ALWPs decreased LPS-induced <t>FAK</t> phosphorylation in BV2 microglial cells. (A) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h, followed by treatment with LPS (1 μg/ml) or PBS for 45 min, and western blotting was performed with <t>anti-p-FAK</t> and anti-FAK antibodies. (B) Quantification of the data from (A) (p-FAK and FAK: con, n = 8; LPS, n = 8; ALWPs + LPS, n = 8). (C) BV2 microglial cells were pretreated with PF-573228 (a FAK inhibitor, 5 μM) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min and finally LPS (1 μg/ml) or PBS for 5 h. Total RNA was isolated, and IL-1β mRNA levels were measured by RT-PCR. (D) Quantification of the data from (C) (con, n = 30; LPS, n = 30; ALWPs + LPS, n = 30; PF-573228 + LPS, n = 30; PF-573228 + ALWPs + LPS, n = 30). ∗∗∗ p
    Rabbit Anti P Fak, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p fak/product/Cell Signaling Technology Inc
    Average 88 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p fak - by Bioz Stars, 2020-09
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    92
    Cell Signaling Technology Inc rabbit anti c cbl
    IFT20 interacts with <t>Cbl-b</t> and c-Cbl, which is targeted to the cilium in PDGF-AA–stimulated cells. (a) Validation of the IFT20-c-Cbl interaction. HEK293T cells coexpressing GFP-IFT20 with either FLAG–tagged WT c-Cbl (FLAG–c-Cbl WT ) or empty FLAG vector were subjected to FLAG IP, followed by WB analysis. (b and c) Reciprocal IPs of endogenous IFT20 or c-Cbl from HEK293T cells. (d) FLAG IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–tagged RING mutant (p.C381A) c-Cbl (FLAG–c-Cbl *RING ). (e) IFT20 IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–c-Cbl *RING , followed by WB analysis. (f) IFM of WT NIH3T3 cells coexpressing GFP-tagged c-Cbl and FLAG–tagged IFT20. The dashed line indicates the Golgi complex, the arrow shows the primary cilium, and the asterisks identify the ciliary base. The nucleus was visualized with DAPI. (g) IFM of WT NIH3T3 cells expressing GFP-tagged c-Cbl. The primary cilium was marked with anti–ARL13B (arrow). The dashed line indicates the Golgi complex, and asterisk indicates the ciliary base. The nucleus was visualized with DAPI. The outline of the cells in panels f and g is highlighted with a dashed line. (h and i) IFM of WT NIH3T3 cells expressing either GFP-tagged c-Cbl (h) or stained with <t>anti–c-Cbl</t> (i). The Golgi complex (dashed line) was labeled with anti–GM130, and the nucleus was visualized with DAPI. (j) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells, labeled with anti–Ac-tub (arrow), in WT NIH3T3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (k) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells and labeled with anti–ARL13B (arrow) in IMCD3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (l) Quantification of relative levels of c-Cbl in the primary cilium shown in panel k. Fluorescence was normalized to background levels. For each of the three experiments, > 15 cells were quantified. Error bars represent means ± SEM ( n = 3).
    Rabbit Anti C Cbl, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti c cbl/product/Cell Signaling Technology Inc
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rabbit anti c cbl - by Bioz Stars, 2020-09
    92/100 stars
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    93
    Cell Signaling Technology Inc rabbit anti human pan 4ebp1
    PRE, W3, and W6 mTORc1 signaling marker differences between LOW and HIGH responders. (A–D) Phosphorylated (p-) mTORc1 targets. There were no significant cluster effects or cluster × time interactions for said markers. p-mTOR and <t>p-4EBP1</t> demonstrated significant time effects. (E–H) Total (pan) mTOR, p70s6k, 4EBP1, and AMPKα. Again, there were no significant cluster effects or cluster × time interactions for said markers, although there were significant time effects for pan mTOR, pan p70s6k, and pan AMPKα. Values presented in line graphs are mean (standard deviation) values. (I) Representative Western blot images for a low and high responder.
    Rabbit Anti Human Pan 4ebp1, supplied by Cell Signaling Technology Inc, 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/rabbit anti human pan 4ebp1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti human pan 4ebp1 - by Bioz Stars, 2020-09
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    92
    Cell Signaling Technology Inc rabbit monoclonal anti pkd antibody
    Effects of <t>PKD</t> inhibition on meiotic spindle configurations and chromosome alignment. ( A ) Representative images of spindle (green) morphologies and chromosome (blue) alignment in the oocytes from control and CID755673 treatment groups. It showed a typical barrel-shaped spindles and well aligned chromosomes in control oocytes; while, unfocused or tripolar poles spindles with agglutinative/scattered chromosomes were successively showed in PKD inhibition groups. Green,α-tubulin; blue, DNA; Bar = 5 μm. ( B ) The percentage of spindle/chromosome defects were recorded in control and CID755673-treated oocytes. ( C ) Protein levels of <t>p-MAPK</t> in control and CID755673 treatment oocytes were determined by western blotting. Data are presented as mean ± s.d. from at least three independent experiments. **, significant difference ( P
    Rabbit Monoclonal Anti Pkd Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti pkd antibody/product/Cell Signaling Technology Inc
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    ALWPs decreased LPS-induced FAK phosphorylation in BV2 microglial cells. (A) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h, followed by treatment with LPS (1 μg/ml) or PBS for 45 min, and western blotting was performed with anti-p-FAK and anti-FAK antibodies. (B) Quantification of the data from (A) (p-FAK and FAK: con, n = 8; LPS, n = 8; ALWPs + LPS, n = 8). (C) BV2 microglial cells were pretreated with PF-573228 (a FAK inhibitor, 5 μM) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min and finally LPS (1 μg/ml) or PBS for 5 h. Total RNA was isolated, and IL-1β mRNA levels were measured by RT-PCR. (D) Quantification of the data from (C) (con, n = 30; LPS, n = 30; ALWPs + LPS, n = 30; PF-573228 + LPS, n = 30; PF-573228 + ALWPs + LPS, n = 30). ∗∗∗ p

    Journal: Frontiers in Aging Neuroscience

    Article Title: An Aqueous Extract of Herbal Medicine ALWPs Enhances Cognitive Performance and Inhibits LPS-Induced Neuroinflammation via FAK/NF-κB Signaling Pathways

    doi: 10.3389/fnagi.2018.00269

    Figure Lengend Snippet: ALWPs decreased LPS-induced FAK phosphorylation in BV2 microglial cells. (A) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h, followed by treatment with LPS (1 μg/ml) or PBS for 45 min, and western blotting was performed with anti-p-FAK and anti-FAK antibodies. (B) Quantification of the data from (A) (p-FAK and FAK: con, n = 8; LPS, n = 8; ALWPs + LPS, n = 8). (C) BV2 microglial cells were pretreated with PF-573228 (a FAK inhibitor, 5 μM) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min and finally LPS (1 μg/ml) or PBS for 5 h. Total RNA was isolated, and IL-1β mRNA levels were measured by RT-PCR. (D) Quantification of the data from (C) (con, n = 30; LPS, n = 30; ALWPs + LPS, n = 30; PF-573228 + LPS, n = 30; PF-573228 + ALWPs + LPS, n = 30). ∗∗∗ p

    Article Snippet: Antibodies and Chemicals We used the following primary antibodies: mouse anti-β-actin (Cat No: sc-47778, Santa Cruz Biotechnology, Dallas, TX, United States), mouse anti-p-IκBα (B-9, Cat No: sc-8404, Santa Cruz Biotechnology), mouse anti-IκBα (Cat No: sc-1643, Santa Cruz Biotechnology), rabbit anti-NF-κB (P65, Cat No: sc-8008, Santa Cruz Biotechnology), mouse anti-PCNA (Cat No: sc-56, Santa Cruz Biotechnology), rat anti-mouse CD11b (M1/70, Cat No: ab8878, Abcam, Cambridge, MA, United States), rabbit anti-FAK (Cat No: 13009S, Cell Signaling Technology, Danvers, MA, United States), rabbit anti-p-FAK (Tyr397, Cat No: 8556S, Cell Signaling Technology), rabbit anti-p-NF-κB (Ser536, Cat No: 3033L, Cell Signaling Technology), rabbit anti-ERK (Cat No: 9102S, Cell Signaling Technology), rabbit anti-p-ERK (Thr202/Tyr204, Cat No:9101S, Cell Signaling Technology), rabbit anti-JNK (Cat No: MBS8509129, MyBioSource, San Diego, CA, United States), rabbit anti-p-JNK (Thr183/Tyr185, Cat No: MBS8508944, MyBioSource), rabbit anti-P38 (Cat No: 9212S, Cell Signaling Technology), rabbit anti-p-P38 (Cat No: 9211S, Cell Signaling Technology), rabbit anti-TLR4 (Cat No: PA5-11597, Thermo Scientific, Waltham, MA, United States), and rabbit anti-TLR4 (Cat No: NB100-56566, Novus Biologicals, Littleton, CO, United States).

    Techniques: Western Blot, Isolation, Reverse Transcription Polymerase Chain Reaction

    ALWPs decreased LPS-induced nuclear NF-κB (Ser536) levels. (A) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h, followed by treatment with LPS (1 μg/ml) or PBS for 45 min and subcellular fractionation (nucleus vs. cytosol). Western blotting was conducted on the cytosolic fraction using antibodies against p-IκBα, IκBα, NF-κB, and β-actin (as a cytosolic marker). (B–D) Quantification of the data from (A) (p-IκBα: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12; IκBα: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12; NF-κB: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12). (E) Western blotting was performed on the nuclear fraction using antibodies against NF-κB and PCNA (as a nuclear marker). (F) Quantification of the data from (E) (con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12). (G) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h and then treated with LPS (1 μg/ml) or PBS for 45 min. Cells were fixed and immunostained with anti-CD11b and anti-p-NF-κB (Ser536) antibodies (40× confocal images). (H) Quantification of the data from (G) (con, n = 122 cells; LPS, n = 129 cells; LPS + ALWPs, n = 111 cells). (I) BV2 microglial cells were pretreated with PF-573228 (a FAK inhibitor, 5 μM) or vehicle (1% DMSO) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min and finally LPS (1 μg/ml) or PBS for 5 h. Cells were fixed and immunostained with anti-CD11b and anti-p-NF-κB (Ser536) antibodies. (J) Quantification of the data from (I) (con, n = 260 cells; LPS, n = 331 cells; LPS + ALWPs, n = 265 cells; LPS + FAK inhibitor, n = 188 cells; LPS + FAK inhibitor + ALWPs, n = 199 cells). Scale bar 20 μm (40× confocal images). ∗ p

    Journal: Frontiers in Aging Neuroscience

    Article Title: An Aqueous Extract of Herbal Medicine ALWPs Enhances Cognitive Performance and Inhibits LPS-Induced Neuroinflammation via FAK/NF-κB Signaling Pathways

    doi: 10.3389/fnagi.2018.00269

    Figure Lengend Snippet: ALWPs decreased LPS-induced nuclear NF-κB (Ser536) levels. (A) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h, followed by treatment with LPS (1 μg/ml) or PBS for 45 min and subcellular fractionation (nucleus vs. cytosol). Western blotting was conducted on the cytosolic fraction using antibodies against p-IκBα, IκBα, NF-κB, and β-actin (as a cytosolic marker). (B–D) Quantification of the data from (A) (p-IκBα: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12; IκBα: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12; NF-κB: con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12). (E) Western blotting was performed on the nuclear fraction using antibodies against NF-κB and PCNA (as a nuclear marker). (F) Quantification of the data from (E) (con, n = 12; LPS, n = 12; ALWPs + LPS, n = 12). (G) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 5 h and then treated with LPS (1 μg/ml) or PBS for 45 min. Cells were fixed and immunostained with anti-CD11b and anti-p-NF-κB (Ser536) antibodies (40× confocal images). (H) Quantification of the data from (G) (con, n = 122 cells; LPS, n = 129 cells; LPS + ALWPs, n = 111 cells). (I) BV2 microglial cells were pretreated with PF-573228 (a FAK inhibitor, 5 μM) or vehicle (1% DMSO) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min and finally LPS (1 μg/ml) or PBS for 5 h. Cells were fixed and immunostained with anti-CD11b and anti-p-NF-κB (Ser536) antibodies. (J) Quantification of the data from (I) (con, n = 260 cells; LPS, n = 331 cells; LPS + ALWPs, n = 265 cells; LPS + FAK inhibitor, n = 188 cells; LPS + FAK inhibitor + ALWPs, n = 199 cells). Scale bar 20 μm (40× confocal images). ∗ p

    Article Snippet: Antibodies and Chemicals We used the following primary antibodies: mouse anti-β-actin (Cat No: sc-47778, Santa Cruz Biotechnology, Dallas, TX, United States), mouse anti-p-IκBα (B-9, Cat No: sc-8404, Santa Cruz Biotechnology), mouse anti-IκBα (Cat No: sc-1643, Santa Cruz Biotechnology), rabbit anti-NF-κB (P65, Cat No: sc-8008, Santa Cruz Biotechnology), mouse anti-PCNA (Cat No: sc-56, Santa Cruz Biotechnology), rat anti-mouse CD11b (M1/70, Cat No: ab8878, Abcam, Cambridge, MA, United States), rabbit anti-FAK (Cat No: 13009S, Cell Signaling Technology, Danvers, MA, United States), rabbit anti-p-FAK (Tyr397, Cat No: 8556S, Cell Signaling Technology), rabbit anti-p-NF-κB (Ser536, Cat No: 3033L, Cell Signaling Technology), rabbit anti-ERK (Cat No: 9102S, Cell Signaling Technology), rabbit anti-p-ERK (Thr202/Tyr204, Cat No:9101S, Cell Signaling Technology), rabbit anti-JNK (Cat No: MBS8509129, MyBioSource, San Diego, CA, United States), rabbit anti-p-JNK (Thr183/Tyr185, Cat No: MBS8508944, MyBioSource), rabbit anti-P38 (Cat No: 9212S, Cell Signaling Technology), rabbit anti-p-P38 (Cat No: 9211S, Cell Signaling Technology), rabbit anti-TLR4 (Cat No: PA5-11597, Thermo Scientific, Waltham, MA, United States), and rabbit anti-TLR4 (Cat No: NB100-56566, Novus Biologicals, Littleton, CO, United States).

    Techniques: Fractionation, Western Blot, Marker

    IFT20 interacts with Cbl-b and c-Cbl, which is targeted to the cilium in PDGF-AA–stimulated cells. (a) Validation of the IFT20-c-Cbl interaction. HEK293T cells coexpressing GFP-IFT20 with either FLAG–tagged WT c-Cbl (FLAG–c-Cbl WT ) or empty FLAG vector were subjected to FLAG IP, followed by WB analysis. (b and c) Reciprocal IPs of endogenous IFT20 or c-Cbl from HEK293T cells. (d) FLAG IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–tagged RING mutant (p.C381A) c-Cbl (FLAG–c-Cbl *RING ). (e) IFT20 IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–c-Cbl *RING , followed by WB analysis. (f) IFM of WT NIH3T3 cells coexpressing GFP-tagged c-Cbl and FLAG–tagged IFT20. The dashed line indicates the Golgi complex, the arrow shows the primary cilium, and the asterisks identify the ciliary base. The nucleus was visualized with DAPI. (g) IFM of WT NIH3T3 cells expressing GFP-tagged c-Cbl. The primary cilium was marked with anti–ARL13B (arrow). The dashed line indicates the Golgi complex, and asterisk indicates the ciliary base. The nucleus was visualized with DAPI. The outline of the cells in panels f and g is highlighted with a dashed line. (h and i) IFM of WT NIH3T3 cells expressing either GFP-tagged c-Cbl (h) or stained with anti–c-Cbl (i). The Golgi complex (dashed line) was labeled with anti–GM130, and the nucleus was visualized with DAPI. (j) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells, labeled with anti–Ac-tub (arrow), in WT NIH3T3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (k) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells and labeled with anti–ARL13B (arrow) in IMCD3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (l) Quantification of relative levels of c-Cbl in the primary cilium shown in panel k. Fluorescence was normalized to background levels. For each of the three experiments, > 15 cells were quantified. Error bars represent means ± SEM ( n = 3).

    Journal: The Journal of Cell Biology

    Article Title: IFT20 modulates ciliary PDGFRα signaling by regulating the stability of Cbl E3 ubiquitin ligases

    doi: 10.1083/jcb.201611050

    Figure Lengend Snippet: IFT20 interacts with Cbl-b and c-Cbl, which is targeted to the cilium in PDGF-AA–stimulated cells. (a) Validation of the IFT20-c-Cbl interaction. HEK293T cells coexpressing GFP-IFT20 with either FLAG–tagged WT c-Cbl (FLAG–c-Cbl WT ) or empty FLAG vector were subjected to FLAG IP, followed by WB analysis. (b and c) Reciprocal IPs of endogenous IFT20 or c-Cbl from HEK293T cells. (d) FLAG IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–tagged RING mutant (p.C381A) c-Cbl (FLAG–c-Cbl *RING ). (e) IFT20 IP of HEK293T cell extracts expressing FLAG–c-Cbl WT or FLAG–c-Cbl *RING , followed by WB analysis. (f) IFM of WT NIH3T3 cells coexpressing GFP-tagged c-Cbl and FLAG–tagged IFT20. The dashed line indicates the Golgi complex, the arrow shows the primary cilium, and the asterisks identify the ciliary base. The nucleus was visualized with DAPI. (g) IFM of WT NIH3T3 cells expressing GFP-tagged c-Cbl. The primary cilium was marked with anti–ARL13B (arrow). The dashed line indicates the Golgi complex, and asterisk indicates the ciliary base. The nucleus was visualized with DAPI. The outline of the cells in panels f and g is highlighted with a dashed line. (h and i) IFM of WT NIH3T3 cells expressing either GFP-tagged c-Cbl (h) or stained with anti–c-Cbl (i). The Golgi complex (dashed line) was labeled with anti–GM130, and the nucleus was visualized with DAPI. (j) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells, labeled with anti–Ac-tub (arrow), in WT NIH3T3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (k) Localization of c-Cbl to the primary cilium in WT NIH3T3 cells and labeled with anti–ARL13B (arrow) in IMCD3 cells before and after stimulation with PDGF-AA. The asterisk marks the ciliary base. The nucleus was visualized with DAPI. (l) Quantification of relative levels of c-Cbl in the primary cilium shown in panel k. Fluorescence was normalized to background levels. For each of the three experiments, > 15 cells were quantified. Error bars represent means ± SEM ( n = 3).

    Article Snippet: Reagents The following antibodies were used: R & D Systems: goat anti-PDGFRα (AF1062); Proteintech: rabbit anti–human IFT20 (13615–1-AP), rabbit anti-IFT88 (13967-1-AP); Sigma-Aldrich: mouse anti–α-tubulin (T5168), mouse anti–acetylated α-tubulin (T6793), mouse anti-FLAG (M2; F1802), rabbit anti-FLAG (F7425); BD Biosciences: mouse anti-CDK1 (610038), mouse anti-p150Glued (610474), mouse anti-GM130 (610823); Enzo life Sciences: mouse anti-giantin (ALX-804-600-C100), mouse anti–mono- and polyubiquitin (FK2; BML-PW8810); Abcam: mouse anti–c-Cbl (Ab119954), rabbit anti–c-Cbl (Ab32027), mouse anti-GFP (Ab1218), chicken anti-GFP (Ab13970), rabbit anti–detyrosinated α-tubulin (Ab48389), rabbit anti-PDGFRα (Ab134123); Cell Signaling Technology: rabbit anti-Myc (2278), rabbit anti-GAPDH (2118), rabbit anti–Cbl-b (9498), rabbit anti-Akt (9272), rabbit anti–phosphorylated Akt (Ser473; 4060), rabbit anti-Erk1/2 (9102), rabbit anti–phosphorylated Erk1/2 (Thr202/Tyr204; 9101), rabbit anti–phosphorylated Rb (9308); Santa Cruz Biotechnology: rabbit anti–c-Cblsc-170), mouse anti–c-Cbl (sc-1651), mouse anti-GFP (sc-9996), rabbit anti-GFP (sc-8334), rabbit anti–phosphorylated PDGFRα (Tyr754; sc-12911R).

    Techniques: Plasmid Preparation, Western Blot, Expressing, Mutagenesis, Staining, Labeling, Fluorescence

    PRE, W3, and W6 mTORc1 signaling marker differences between LOW and HIGH responders. (A–D) Phosphorylated (p-) mTORc1 targets. There were no significant cluster effects or cluster × time interactions for said markers. p-mTOR and p-4EBP1 demonstrated significant time effects. (E–H) Total (pan) mTOR, p70s6k, 4EBP1, and AMPKα. Again, there were no significant cluster effects or cluster × time interactions for said markers, although there were significant time effects for pan mTOR, pan p70s6k, and pan AMPKα. Values presented in line graphs are mean (standard deviation) values. (I) Representative Western blot images for a low and high responder.

    Journal: Frontiers in Physiology

    Article Title: Pre-training Skeletal Muscle Fiber Size and Predominant Fiber Type Best Predict Hypertrophic Responses to 6 Weeks of Resistance Training in Previously Trained Young Men

    doi: 10.3389/fphys.2019.00297

    Figure Lengend Snippet: PRE, W3, and W6 mTORc1 signaling marker differences between LOW and HIGH responders. (A–D) Phosphorylated (p-) mTORc1 targets. There were no significant cluster effects or cluster × time interactions for said markers. p-mTOR and p-4EBP1 demonstrated significant time effects. (E–H) Total (pan) mTOR, p70s6k, 4EBP1, and AMPKα. Again, there were no significant cluster effects or cluster × time interactions for said markers, although there were significant time effects for pan mTOR, pan p70s6k, and pan AMPKα. Values presented in line graphs are mean (standard deviation) values. (I) Representative Western blot images for a low and high responder.

    Article Snippet: Rabbit anti-human phospho-p70s6k (Thr389) (1:1,000; catalog #: 9234; Cell Signaling), rabbit anti-human pan p70s6k (1:1,000; catalog #: 2708; Cell Signaling), rabbit anti-human phospho-4EBP1 (Thr37/46) (1:1,000; catalog #: 2855; Cell Signaling), rabbit anti-human pan 4EBP1 (1:1,000; catalog #: 9644; Cell Signaling), rabbit anti-human phospho-mTOR (Ser2448) (1:1,000; catalog #: 2971; Cell Signaling), rabbit anti-human pan mTOR (1:1,000; catalog #: 2972; Cell Signaling), rabbit anti-human phospho-AMPKα (Thr172) (1:1,000; catalog #: 2535; Cell Signaling), rabbit anti-human pan AMPKα (1:1,000; catalog #: 2532; Cell Signaling), rabbit anti-human androgen receptor (1:1,000; catalog #: 5153; Cell Signaling) and rabbit anti-human ubiquitin (1:1,000; catalog #: 3933; Cell Signaling) were incubated with membranes overnight at 4°C in TBST with 5% bovine serum albumin (BSA).

    Techniques: Marker, Standard Deviation, Western Blot

    Effects of PKD inhibition on meiotic spindle configurations and chromosome alignment. ( A ) Representative images of spindle (green) morphologies and chromosome (blue) alignment in the oocytes from control and CID755673 treatment groups. It showed a typical barrel-shaped spindles and well aligned chromosomes in control oocytes; while, unfocused or tripolar poles spindles with agglutinative/scattered chromosomes were successively showed in PKD inhibition groups. Green,α-tubulin; blue, DNA; Bar = 5 μm. ( B ) The percentage of spindle/chromosome defects were recorded in control and CID755673-treated oocytes. ( C ) Protein levels of p-MAPK in control and CID755673 treatment oocytes were determined by western blotting. Data are presented as mean ± s.d. from at least three independent experiments. **, significant difference ( P

    Journal: Aging (Albany NY)

    Article Title: Inhibition of protein kinase D disrupts spindle formation and actin assembly during porcine oocyte maturation

    doi: 10.18632/aging.101667

    Figure Lengend Snippet: Effects of PKD inhibition on meiotic spindle configurations and chromosome alignment. ( A ) Representative images of spindle (green) morphologies and chromosome (blue) alignment in the oocytes from control and CID755673 treatment groups. It showed a typical barrel-shaped spindles and well aligned chromosomes in control oocytes; while, unfocused or tripolar poles spindles with agglutinative/scattered chromosomes were successively showed in PKD inhibition groups. Green,α-tubulin; blue, DNA; Bar = 5 μm. ( B ) The percentage of spindle/chromosome defects were recorded in control and CID755673-treated oocytes. ( C ) Protein levels of p-MAPK in control and CID755673 treatment oocytes were determined by western blotting. Data are presented as mean ± s.d. from at least three independent experiments. **, significant difference ( P

    Article Snippet: Rabbit monoclonal anti-PKD antibody (#2052), rabbit polyclonal anti-phospho-MAPK antibody (#4370), rabbit monoclonal anti-p-cofilin antibody (#3313), mouse monoclonal anti-β-actin antibody (#3700), rabbit monoclonal anti-GAPDH antibody (#2118) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Inhibition, Western Blot