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    Santa Cruz Biotechnology gapdh
    Overexpression of SOX9 transactivates NEDD9 and induces MMPs expression. a qPCR analysis for the expression levels of <t>SOX10</t> , SOX9 , and NEDD9 in A375M and WM266–4 cell lines treated with scramble control, SOX10 KD alone and SOX10 KD together with increasing amount of SOX9 OE lentiviruses. b Immunoblotting for the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with the indicated constructs. The intensity of protein bands in arbitrary units for SOX10, SOX9, and NEDD9 in each melanoma cell line is relative to scramble control which is set to 1 as a reference. The red arrow indicates the phosphorylated form of NEDD9. Asterisk indicates non-specific bands. <t>GAPDH</t> serves as a loading control. c A375M and WM266–4 cells were transfected with a 1 kb-NEDD9 promoter-driven luciferase reporter construct plus renilla for normalization of transfection efficiency together with scramble control, SOX10 KD, SOX10 KD plus increasing amount of SOX9 OE lentiviruses, SOX10 OE, and SOX9 OE. Fold activation of three independent luciferase assays. SOX10 KD is set to 1 as a reference. d Schematic diagram showing the presence of a SOX binding motif within the 167 bp DNA fragment detected by ChIP-qPCR whereas the 284 bp fragment serves as a negative control. ChIP-qPCR data showing a higher DNA binding capacity by SOX10 than SOX9. Anti-IgG serves as a negative control. e Western blot analysis using the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with vehicle control and SOX9 OE. The red arrow indicates the phosphorylated form of NEDD9. f DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. g Transwell invasion assay for each cell line treated with the vehicle alone and SOX9 OE lentivirus. Scale bars: 100 μM. h mRNA expression of SOX9 and members of MMP family were quantified by qRT-PCR in A375M and WM266–4 cells treated with the vehicle alone and SOX9 OE. Error bars represent mean ± SD of three independent experiments. n.s, non-significant; * p
    Gapdh, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gapdh/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gapdh - by Bioz Stars, 2021-03
    86/100 stars
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    gapdh  (Abcam)
    99
    Abcam gapdh
    Effect of the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) on the nuclear factor (NF)-κB signaling pathway in xanthohumol (Xn)-treated AGS cells. Cells were pre-treated with the ROS inhibitor NAC (5 mM) for 1 h, and then treated with Xn (20 µ M) for 24 h; they were then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Expression of IκBα and p-IκBα protein; (D-F) Expression of nuclear and cytosolic p65 protein. Histone <t>H3</t> served as the nuclear loading control, <t>GAPDH</t> served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. **P
    Gapdh, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gapdh/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gapdh - by Bioz Stars, 2021-03
    99/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc anti gapdh
    Effect of the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) on the nuclear factor (NF)-κB signaling pathway in xanthohumol (Xn)-treated AGS cells. Cells were pre-treated with the ROS inhibitor NAC (5 mM) for 1 h, and then treated with Xn (20 µ M) for 24 h; they were then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Expression of IκBα and p-IκBα protein; (D-F) Expression of nuclear and cytosolic p65 protein. Histone <t>H3</t> served as the nuclear loading control, <t>GAPDH</t> served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. **P
    Anti Gapdh, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti gapdh - by Bioz Stars, 2021-03
    99/100 stars
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    N/A
    Rabbit polyclonal antibody to Attachment protein G3P Isotype Note IgG Host Note Rabbit Conjugation Note Unconjugated Reactivity Note Viral Application Note ELISA
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    Image Search Results


    Overexpression of SOX9 transactivates NEDD9 and induces MMPs expression. a qPCR analysis for the expression levels of SOX10 , SOX9 , and NEDD9 in A375M and WM266–4 cell lines treated with scramble control, SOX10 KD alone and SOX10 KD together with increasing amount of SOX9 OE lentiviruses. b Immunoblotting for the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with the indicated constructs. The intensity of protein bands in arbitrary units for SOX10, SOX9, and NEDD9 in each melanoma cell line is relative to scramble control which is set to 1 as a reference. The red arrow indicates the phosphorylated form of NEDD9. Asterisk indicates non-specific bands. GAPDH serves as a loading control. c A375M and WM266–4 cells were transfected with a 1 kb-NEDD9 promoter-driven luciferase reporter construct plus renilla for normalization of transfection efficiency together with scramble control, SOX10 KD, SOX10 KD plus increasing amount of SOX9 OE lentiviruses, SOX10 OE, and SOX9 OE. Fold activation of three independent luciferase assays. SOX10 KD is set to 1 as a reference. d Schematic diagram showing the presence of a SOX binding motif within the 167 bp DNA fragment detected by ChIP-qPCR whereas the 284 bp fragment serves as a negative control. ChIP-qPCR data showing a higher DNA binding capacity by SOX10 than SOX9. Anti-IgG serves as a negative control. e Western blot analysis using the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with vehicle control and SOX9 OE. The red arrow indicates the phosphorylated form of NEDD9. f DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. g Transwell invasion assay for each cell line treated with the vehicle alone and SOX9 OE lentivirus. Scale bars: 100 μM. h mRNA expression of SOX9 and members of MMP family were quantified by qRT-PCR in A375M and WM266–4 cells treated with the vehicle alone and SOX9 OE. Error bars represent mean ± SD of three independent experiments. n.s, non-significant; * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: SOX9 is a dose-dependent metastatic fate determinant in melanoma

    doi: 10.1186/s13046-018-0998-6

    Figure Lengend Snippet: Overexpression of SOX9 transactivates NEDD9 and induces MMPs expression. a qPCR analysis for the expression levels of SOX10 , SOX9 , and NEDD9 in A375M and WM266–4 cell lines treated with scramble control, SOX10 KD alone and SOX10 KD together with increasing amount of SOX9 OE lentiviruses. b Immunoblotting for the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with the indicated constructs. The intensity of protein bands in arbitrary units for SOX10, SOX9, and NEDD9 in each melanoma cell line is relative to scramble control which is set to 1 as a reference. The red arrow indicates the phosphorylated form of NEDD9. Asterisk indicates non-specific bands. GAPDH serves as a loading control. c A375M and WM266–4 cells were transfected with a 1 kb-NEDD9 promoter-driven luciferase reporter construct plus renilla for normalization of transfection efficiency together with scramble control, SOX10 KD, SOX10 KD plus increasing amount of SOX9 OE lentiviruses, SOX10 OE, and SOX9 OE. Fold activation of three independent luciferase assays. SOX10 KD is set to 1 as a reference. d Schematic diagram showing the presence of a SOX binding motif within the 167 bp DNA fragment detected by ChIP-qPCR whereas the 284 bp fragment serves as a negative control. ChIP-qPCR data showing a higher DNA binding capacity by SOX10 than SOX9. Anti-IgG serves as a negative control. e Western blot analysis using the indicated antibodies on protein lysates derived from A375M and WM266–4 cells treated with vehicle control and SOX9 OE. The red arrow indicates the phosphorylated form of NEDD9. f DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. g Transwell invasion assay for each cell line treated with the vehicle alone and SOX9 OE lentivirus. Scale bars: 100 μM. h mRNA expression of SOX9 and members of MMP family were quantified by qRT-PCR in A375M and WM266–4 cells treated with the vehicle alone and SOX9 OE. Error bars represent mean ± SD of three independent experiments. n.s, non-significant; * p

    Article Snippet: Membranes were probed with antibodies against SOX9 (H-90, Santa Cruz), SOX10 (N-20, Santa Cruz), NEDD9 (Clone 2G9, Abcam) and GAPDH (FL-335, Santa Cruz) for overnight at 4 °C and then incubated with appropriate horseradish peroxidase-conjugated goat anti-rabbit, rabbit anti-mouse and donkey anti-goat antibodies (at 1:2000, Dako) at room temperature for 1 h. After incubation with ECL substrate (WesternBright, Advansta) for 1–3 min, blots were exposed to X-ray film (FujiFilm Super RX) at different times to obtain the optimal intensity of the protein bands which were analyzed by ImageJ.

    Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, Construct, Transfection, Luciferase, Activation Assay, Binding Assay, Chromatin Immunoprecipitation, Negative Control, Western Blot, Transwell Invasion Assay, Quantitative RT-PCR

    SOXE directs melanoma mesenchymal migration through the NEDD9-mediated focal adhesion dynamics and RHO GTPase signaling. a A375M cells treated with the indicated constructs were stained for vinculin and phalloidin. Cell nuclei were counterstained with DAPI. Scale bar: 50 μM. The number of vinculin per cell ( b ), area of vinculin per cell ( c ) and the average size of single vinculin per cell ( d ) were quantified. Thirty cells were analyzed for each treatment. e A375M cells treated with the indicated constructs were subjected to RHOA and RAC1 activation assays. GAPDH serves as a loading control. f Quantification of band intensity from the densitometric analysis. g Schematic model showing a dose-dependent role of SOX9 within a heterogeneous population of melanoma in which low level of SOX9 expression is anti-tumorigenic and high SOX9 is oncogenic. Scale bar: 100 μM. Error bars represent ± SD of three independent experiments. n.s. non-significant, * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: SOX9 is a dose-dependent metastatic fate determinant in melanoma

    doi: 10.1186/s13046-018-0998-6

    Figure Lengend Snippet: SOXE directs melanoma mesenchymal migration through the NEDD9-mediated focal adhesion dynamics and RHO GTPase signaling. a A375M cells treated with the indicated constructs were stained for vinculin and phalloidin. Cell nuclei were counterstained with DAPI. Scale bar: 50 μM. The number of vinculin per cell ( b ), area of vinculin per cell ( c ) and the average size of single vinculin per cell ( d ) were quantified. Thirty cells were analyzed for each treatment. e A375M cells treated with the indicated constructs were subjected to RHOA and RAC1 activation assays. GAPDH serves as a loading control. f Quantification of band intensity from the densitometric analysis. g Schematic model showing a dose-dependent role of SOX9 within a heterogeneous population of melanoma in which low level of SOX9 expression is anti-tumorigenic and high SOX9 is oncogenic. Scale bar: 100 μM. Error bars represent ± SD of three independent experiments. n.s. non-significant, * p

    Article Snippet: Membranes were probed with antibodies against SOX9 (H-90, Santa Cruz), SOX10 (N-20, Santa Cruz), NEDD9 (Clone 2G9, Abcam) and GAPDH (FL-335, Santa Cruz) for overnight at 4 °C and then incubated with appropriate horseradish peroxidase-conjugated goat anti-rabbit, rabbit anti-mouse and donkey anti-goat antibodies (at 1:2000, Dako) at room temperature for 1 h. After incubation with ECL substrate (WesternBright, Advansta) for 1–3 min, blots were exposed to X-ray film (FujiFilm Super RX) at different times to obtain the optimal intensity of the protein bands which were analyzed by ImageJ.

    Techniques: Migration, Construct, Staining, Activation Assay, Expressing

    Overexpression of SOX9 and NEDD9 restore the oncogenic features of SOX10 KD melanoma cells. qRT-PCR ( a ) and Western blot ( b ) analysis for the expression levels of SOX10, SOX9 and NEDD9 in A375M and WM266–4 cell lines treated with the indicated constructs. Data are fold change normalized to scramble control and the average of three independent assays. The red arrow indicates the phosphorylated form of NEDD9. GAPDH serves as a loading control. AlamarBlue ( c ) and transwell invasion assays ( d ) of each cell line treated with the indicated constructs. e DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. Scale bars: 100 μM. f Representative images of crystal violet stained A375M and WM266–4 clones subjected to different treatments. g Quantification of the number of A375M and WM266–4 colonies treated with the indicated constructs. Error bars represent mean ± SD of three independent experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: SOX9 is a dose-dependent metastatic fate determinant in melanoma

    doi: 10.1186/s13046-018-0998-6

    Figure Lengend Snippet: Overexpression of SOX9 and NEDD9 restore the oncogenic features of SOX10 KD melanoma cells. qRT-PCR ( a ) and Western blot ( b ) analysis for the expression levels of SOX10, SOX9 and NEDD9 in A375M and WM266–4 cell lines treated with the indicated constructs. Data are fold change normalized to scramble control and the average of three independent assays. The red arrow indicates the phosphorylated form of NEDD9. GAPDH serves as a loading control. AlamarBlue ( c ) and transwell invasion assays ( d ) of each cell line treated with the indicated constructs. e DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. Scale bars: 100 μM. f Representative images of crystal violet stained A375M and WM266–4 clones subjected to different treatments. g Quantification of the number of A375M and WM266–4 colonies treated with the indicated constructs. Error bars represent mean ± SD of three independent experiments. * p

    Article Snippet: Membranes were probed with antibodies against SOX9 (H-90, Santa Cruz), SOX10 (N-20, Santa Cruz), NEDD9 (Clone 2G9, Abcam) and GAPDH (FL-335, Santa Cruz) for overnight at 4 °C and then incubated with appropriate horseradish peroxidase-conjugated goat anti-rabbit, rabbit anti-mouse and donkey anti-goat antibodies (at 1:2000, Dako) at room temperature for 1 h. After incubation with ECL substrate (WesternBright, Advansta) for 1–3 min, blots were exposed to X-ray film (FujiFilm Super RX) at different times to obtain the optimal intensity of the protein bands which were analyzed by ImageJ.

    Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Construct, Staining, Clone Assay

    Upregulated or low level of SOX9 expression contributes to the anti-metastatic/anti-oncogenic activities of SOX10 knockdown (KD) melanoma cells. a Expression of SOX10, SOX9, and NEDD9 in human melanocytes (HEMa-LP), and a panel of metastatic melanoma cell lines. GAPDH was used as a loading control. The yellow box indicates protein bands corresponding to the size of SOX9. The red arrow indicates a phosphorylated form of NEDD9. The intensity of protein bands in arbitrary units for SOX10, SOX9, and NEDD9 in each melanoma cell line is relative to HEMa-LP which is set to 1 as a reference. b Line plots represent the intensity of protein bands shown in ( a ). c qRT-PCR analysis of SOX10 , SOX9 and NEDD9 transcript levels in A375M and WM266–4 cells treated with scramble control, SOX10 KD and NEDD9 KD. Data represent fold change normalized to scramble control and the average of three independent assays. d Western blot analysis of SOX9, SOX10 and NEDD9 protein levels in each cell line transduced with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. GAPDH serves as a loading control. The red arrow indicates a phosphorylated form of NEDD9. AlamarBlue ( e ), transwell invasion ( f , g ) and colony formation assays ( h , i ) of each cell line treated with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. g DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. Scale bars: 100 μM ( i ) Representative images showing crystal violet stained colonies formed from A375M and WM266–4 cells treated with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. j Western blot analysis of SOX10, SOX9 and p21 protein levels in each cell line transduced with scramble control, SOX10 KD alone, SOX10 KD + SOX9 KD, two different volume (100 μL, 200 μL) of lentiviruses encoding SOX9 gene ( SOX9 OE) in SOX10 KD and maximum dose of SOX9 OE in parental cells. GAPDH serves as a loading control. Error bars represent mean ± SD of three independent experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: SOX9 is a dose-dependent metastatic fate determinant in melanoma

    doi: 10.1186/s13046-018-0998-6

    Figure Lengend Snippet: Upregulated or low level of SOX9 expression contributes to the anti-metastatic/anti-oncogenic activities of SOX10 knockdown (KD) melanoma cells. a Expression of SOX10, SOX9, and NEDD9 in human melanocytes (HEMa-LP), and a panel of metastatic melanoma cell lines. GAPDH was used as a loading control. The yellow box indicates protein bands corresponding to the size of SOX9. The red arrow indicates a phosphorylated form of NEDD9. The intensity of protein bands in arbitrary units for SOX10, SOX9, and NEDD9 in each melanoma cell line is relative to HEMa-LP which is set to 1 as a reference. b Line plots represent the intensity of protein bands shown in ( a ). c qRT-PCR analysis of SOX10 , SOX9 and NEDD9 transcript levels in A375M and WM266–4 cells treated with scramble control, SOX10 KD and NEDD9 KD. Data represent fold change normalized to scramble control and the average of three independent assays. d Western blot analysis of SOX9, SOX10 and NEDD9 protein levels in each cell line transduced with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. GAPDH serves as a loading control. The red arrow indicates a phosphorylated form of NEDD9. AlamarBlue ( e ), transwell invasion ( f , g ) and colony formation assays ( h , i ) of each cell line treated with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. g DAPI images of transwell invasion of melanoma cells treated with the indicated constructs. Scale bars: 100 μM ( i ) Representative images showing crystal violet stained colonies formed from A375M and WM266–4 cells treated with scramble control, SOX10 KD and SOX10 KD + SOX9 KD. j Western blot analysis of SOX10, SOX9 and p21 protein levels in each cell line transduced with scramble control, SOX10 KD alone, SOX10 KD + SOX9 KD, two different volume (100 μL, 200 μL) of lentiviruses encoding SOX9 gene ( SOX9 OE) in SOX10 KD and maximum dose of SOX9 OE in parental cells. GAPDH serves as a loading control. Error bars represent mean ± SD of three independent experiments. * p

    Article Snippet: Membranes were probed with antibodies against SOX9 (H-90, Santa Cruz), SOX10 (N-20, Santa Cruz), NEDD9 (Clone 2G9, Abcam) and GAPDH (FL-335, Santa Cruz) for overnight at 4 °C and then incubated with appropriate horseradish peroxidase-conjugated goat anti-rabbit, rabbit anti-mouse and donkey anti-goat antibodies (at 1:2000, Dako) at room temperature for 1 h. After incubation with ECL substrate (WesternBright, Advansta) for 1–3 min, blots were exposed to X-ray film (FujiFilm Super RX) at different times to obtain the optimal intensity of the protein bands which were analyzed by ImageJ.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transduction, Construct, Staining

    PKCε protein levels in HeLaPKCεA/E and glioblastoma cell lines. Protein extracts from HeLaPKCεA/E (−Dox), HeLaPKCεA/E (+Dox), U-118 MG, U-138 MG, T98G, LN18 cell lines were subjected to Western blot analysis using antibodies against PKCε and GAPDH. HeLaPKCεA/E cells with doxycycline-induced expression of PKCε (+Dox) and without doxycycline treatment (−Dox) served as a reference cell lines with low and high expression of PKCε, respectively. The average relative absorbance of three independent experiments is presented in a bar graph

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: PKCε protein levels in HeLaPKCεA/E and glioblastoma cell lines. Protein extracts from HeLaPKCεA/E (−Dox), HeLaPKCεA/E (+Dox), U-118 MG, U-138 MG, T98G, LN18 cell lines were subjected to Western blot analysis using antibodies against PKCε and GAPDH. HeLaPKCεA/E cells with doxycycline-induced expression of PKCε (+Dox) and without doxycycline treatment (−Dox) served as a reference cell lines with low and high expression of PKCε, respectively. The average relative absorbance of three independent experiments is presented in a bar graph

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Western Blot, Expressing

    Knockdown of PKCɛ diminishes the level and phosphorylation of Akt in glioma cells. a U-138 MG and b U-118 MG were transfected for 72 h with PKCε siRNA and Control siRNA and then were treated for 24 h with rapamycin (300 nM) or 3-MA (5 mM). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Knockdown of PKCɛ diminishes the level and phosphorylation of Akt in glioma cells. a U-138 MG and b U-118 MG were transfected for 72 h with PKCε siRNA and Control siRNA and then were treated for 24 h with rapamycin (300 nM) or 3-MA (5 mM). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Transfection

    Influence of PKCε downregulation and rapamycin or 3-MA treatment on protein level directly involved in autophagy activation (mTOR, PI3K, Beclin1). a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. In case of mTOR the order of bands has been changing, due to a different final construct of probe layout. The densitometric analysis represents means ±SD of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Influence of PKCε downregulation and rapamycin or 3-MA treatment on protein level directly involved in autophagy activation (mTOR, PI3K, Beclin1). a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. In case of mTOR the order of bands has been changing, due to a different final construct of probe layout. The densitometric analysis represents means ±SD of three independent experiments. * P

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Activation Assay, Transfection, Construct

    Effect of PKCε downregulation and rapamycin or 3-MA treatment on proteins engaged in the formation of a double-membrane structure known as autophagosome (Atg5, LC3-II). a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Effect of PKCε downregulation and rapamycin or 3-MA treatment on proteins engaged in the formation of a double-membrane structure known as autophagosome (Atg5, LC3-II). a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Transfection

    Effect of PKCε downregulation and rapamycin or 3-MA treatment on Bcl-2, a protein that regulates autophagy process. a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Effect of PKCε downregulation and rapamycin or 3-MA treatment on Bcl-2, a protein that regulates autophagy process. a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM) (PKCε siRNA + Rap) or 3-MA (5 mM) (PKCε siRNA + 3-MA). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Transfection

    Effect of PKCε downregulation on the adhesion of glioblastoma cells. a . U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and Control siRNA and then were seeded in culture plates coated with Matrigel (Corning Life Sciences, NY, USA). The photos represent cell adhesion under the microscope at 100 x magnification field. c Evaluation of FAK expression in U-138 MG and U-118 MG cells with knockdown PKCε. Total protein expression and FAK phosphorylation at Tyr-397/Tyr-576/577 were assessed. GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Effect of PKCε downregulation on the adhesion of glioblastoma cells. a . U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and Control siRNA and then were seeded in culture plates coated with Matrigel (Corning Life Sciences, NY, USA). The photos represent cell adhesion under the microscope at 100 x magnification field. c Evaluation of FAK expression in U-138 MG and U-118 MG cells with knockdown PKCε. Total protein expression and FAK phosphorylation at Tyr-397/Tyr-576/577 were assessed. GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Transfection, Microscopy, Expressing

    Modulation SQSTM1/p62 after PRKCE silencing and rapamycin treatment. a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

    doi: 10.1186/s12885-018-4095-1

    Figure Lengend Snippet: Modulation SQSTM1/p62 after PRKCE silencing and rapamycin treatment. a U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and non-targeting siRNA (Control siRNA) and then were treated for 24 h with rapamycin (300 nM). GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P

    Article Snippet: The following antibodies were used for detection: anti-PKCε, anti-BECN1, anti-Bcl-2, anti-FAK, anti-pFAK (Tyr-397), anti-pFAK (Tyr 576/577), anti-GAPDH, anti-rabbit IgG-HRP (all from Santa Cruz Biotechnology, CA, USA), anti-MAPLC3β (Sigma, St Louis, MO, USA), anti-ATG5, anti-mTOR, anti-SQSTM1/p62, anti-Akt and anti-pAkt (Ser473) (from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Transfection

    Effect of the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) on the nuclear factor (NF)-κB signaling pathway in xanthohumol (Xn)-treated AGS cells. Cells were pre-treated with the ROS inhibitor NAC (5 mM) for 1 h, and then treated with Xn (20 µ M) for 24 h; they were then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Expression of IκBα and p-IκBα protein; (D-F) Expression of nuclear and cytosolic p65 protein. Histone H3 served as the nuclear loading control, GAPDH served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. **P

    Journal: Oncology Reports

    Article Title: Xanthohumol, a prenylated flavonoid from Hops, exerts anticancer effects against gastric cancer in vitro

    doi: 10.3892/or.2018.6723

    Figure Lengend Snippet: Effect of the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) on the nuclear factor (NF)-κB signaling pathway in xanthohumol (Xn)-treated AGS cells. Cells were pre-treated with the ROS inhibitor NAC (5 mM) for 1 h, and then treated with Xn (20 µ M) for 24 h; they were then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Expression of IκBα and p-IκBα protein; (D-F) Expression of nuclear and cytosolic p65 protein. Histone H3 served as the nuclear loading control, GAPDH served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. **P

    Article Snippet: Antibodies against Bcl-2 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab194583), Bax (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab32503), p-IκBα (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab133462), IκBα (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab32518), p65 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab16502), histone H3 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab1791) and GAPDH (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab9485) were obtained from Abcam (Cambridge, UK).

    Techniques: Western Blot, Expressing

    Effect of xanthohumol (Xn) on the nuclear factor (NF)-κB signaling pathway in AGS cells. Cells were treated with different concentrations of Xn (0–20 µ M) for 24 h, then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Effects of Xn on IκBα and p-IκBα expression. (D-F) Effect of Xn on nuclear and cytosolic p65 expression. Histone H3 served as the nuclear loading control and GAPDH served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. *P

    Journal: Oncology Reports

    Article Title: Xanthohumol, a prenylated flavonoid from Hops, exerts anticancer effects against gastric cancer in vitro

    doi: 10.3892/or.2018.6723

    Figure Lengend Snippet: Effect of xanthohumol (Xn) on the nuclear factor (NF)-κB signaling pathway in AGS cells. Cells were treated with different concentrations of Xn (0–20 µ M) for 24 h, then harvested and lysed to measure NF-κB signaling proteins through western blotting. (A-C) Effects of Xn on IκBα and p-IκBα expression. (D-F) Effect of Xn on nuclear and cytosolic p65 expression. Histone H3 served as the nuclear loading control and GAPDH served as the cytosolic loading control. Data are expressed as mean ± standard error of the mean. n=3. *P

    Article Snippet: Antibodies against Bcl-2 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab194583), Bax (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab32503), p-IκBα (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab133462), IκBα (rabbit monoclonal antibody, dilution 1:1,000; cat. no. ab32518), p65 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab16502), histone H3 (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab1791) and GAPDH (rabbit polyclonal antibody, dilution 1:1,000; cat. no. ab9485) were obtained from Abcam (Cambridge, UK).

    Techniques: Western Blot, Expressing