rabbit anti sox9 monoclonal antibody  (Abcam)

 
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    Name:
    Goat Anti Rabbit IgG H L HRP
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    AB205718
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    Structured Review

    Abcam rabbit anti sox9 monoclonal antibody
    Down-regulation of <t>SOX9</t> inhibited the growth of tumor in vivo a the tumor specimen figure of nude mice. b statistics of tumor volume, the growth of si-MALAT1 group and si- SOX9 group was the slowest, and the volume was the smallest. The tumor volume of NC group and si- SOX9 + miR-145 inhibitor group, si-MALAT1 + pCDNA3- SOX9 group was comparative. pcDNA3.1- SOX9 group had the largest tumor volume. c Statistical results showed that tumor weight in nude mice, the tumor weight was significantly reduced in si-MALAT1 group. d The relative miR-145 expression was conspicuously increased in si-MALAT1 group in tumor tissues. e , f The qRT-PCR and western blot showed SOX9 expression was significantly decreased in si-MALAT1 group in tumor tissues. ** P

    https://www.bioz.com/result/rabbit anti sox9 monoclonal antibody/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti sox9 monoclonal antibody - by Bioz Stars, 2021-09
    99/100 stars

    Images

    1) Product Images from "The effects of lncRNA MALAT1 on proliferation, invasion and migration in colorectal cancer through regulating SOX9"

    Article Title: The effects of lncRNA MALAT1 on proliferation, invasion and migration in colorectal cancer through regulating SOX9

    Journal: Molecular Medicine

    doi: 10.1186/s10020-018-0050-5

    Down-regulation of SOX9 inhibited the growth of tumor in vivo a the tumor specimen figure of nude mice. b statistics of tumor volume, the growth of si-MALAT1 group and si- SOX9 group was the slowest, and the volume was the smallest. The tumor volume of NC group and si- SOX9 + miR-145 inhibitor group, si-MALAT1 + pCDNA3- SOX9 group was comparative. pcDNA3.1- SOX9 group had the largest tumor volume. c Statistical results showed that tumor weight in nude mice, the tumor weight was significantly reduced in si-MALAT1 group. d The relative miR-145 expression was conspicuously increased in si-MALAT1 group in tumor tissues. e , f The qRT-PCR and western blot showed SOX9 expression was significantly decreased in si-MALAT1 group in tumor tissues. ** P
    Figure Legend Snippet: Down-regulation of SOX9 inhibited the growth of tumor in vivo a the tumor specimen figure of nude mice. b statistics of tumor volume, the growth of si-MALAT1 group and si- SOX9 group was the slowest, and the volume was the smallest. The tumor volume of NC group and si- SOX9 + miR-145 inhibitor group, si-MALAT1 + pCDNA3- SOX9 group was comparative. pcDNA3.1- SOX9 group had the largest tumor volume. c Statistical results showed that tumor weight in nude mice, the tumor weight was significantly reduced in si-MALAT1 group. d The relative miR-145 expression was conspicuously increased in si-MALAT1 group in tumor tissues. e , f The qRT-PCR and western blot showed SOX9 expression was significantly decreased in si-MALAT1 group in tumor tissues. ** P

    Techniques Used: In Vivo, Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot

    Effects of SOX9 on cycle and apoptosis of colorectal cancer cells a the cell cycle was assessed by flow cytometry, the number of cells in G1 phase was the most in the si- SOX9 group and was the least in pcDNA3.1- SOX9 group, the si- SOX9 + miR-145 inhibitor group was comparative to the NC group. b The apoptosis rate which measured by flow cytometry of pcDNA3.1- SOX9 group was the highest, si- SOX9 + miR-145 inhibitor group and NC group took the second place, and si- SOX9 group had the lowest apoptosis rate. * P
    Figure Legend Snippet: Effects of SOX9 on cycle and apoptosis of colorectal cancer cells a the cell cycle was assessed by flow cytometry, the number of cells in G1 phase was the most in the si- SOX9 group and was the least in pcDNA3.1- SOX9 group, the si- SOX9 + miR-145 inhibitor group was comparative to the NC group. b The apoptosis rate which measured by flow cytometry of pcDNA3.1- SOX9 group was the highest, si- SOX9 + miR-145 inhibitor group and NC group took the second place, and si- SOX9 group had the lowest apoptosis rate. * P

    Techniques Used: Flow Cytometry, Cytometry

    Effects of SOX9 on proliferation, invasion and migration of colorectal cancer cells a and b the cell proliferation of colorectal cancer cell lines DLD-1 and HT-29 transfected with different interference sequences, cell proliferation was the weakest in si- SOX9 group, and pcDNA3.1- SOX9 group had the strongest cell proliferation ability compared with NC group. The cell proliferation of si- SOX9 + miR-145 inhibitor group and NC group was comparative. c cell colony formation of colorectal cancer, the formation rate of colorectal cancer cells in the si- SOX9 group was remarkably slower than that of the NC group. The cell formation rate was significantly higher in pcDNA3.1- SOX9 group than that of NC group, and the cell formation in si- SOX9 + miR-145 inhibitor group and NC group had no significant difference. d invasion assays of two kinds of cancer cells under different interference conditions. e the scratch wound healing assays of two kinds of cancer cells under different interference conditions. ** P
    Figure Legend Snippet: Effects of SOX9 on proliferation, invasion and migration of colorectal cancer cells a and b the cell proliferation of colorectal cancer cell lines DLD-1 and HT-29 transfected with different interference sequences, cell proliferation was the weakest in si- SOX9 group, and pcDNA3.1- SOX9 group had the strongest cell proliferation ability compared with NC group. The cell proliferation of si- SOX9 + miR-145 inhibitor group and NC group was comparative. c cell colony formation of colorectal cancer, the formation rate of colorectal cancer cells in the si- SOX9 group was remarkably slower than that of the NC group. The cell formation rate was significantly higher in pcDNA3.1- SOX9 group than that of NC group, and the cell formation in si- SOX9 + miR-145 inhibitor group and NC group had no significant difference. d invasion assays of two kinds of cancer cells under different interference conditions. e the scratch wound healing assays of two kinds of cancer cells under different interference conditions. ** P

    Techniques Used: Migration, Transfection

    SOX9 was the target gene of miR-145. a the binding sites of miR-145 and SOX9 . b results of SOX9 enrichment, SOX9 was the target gene of miR-145. c Transfection efficiency test after the transfection of si- SOX9 or pcDNA3.1- SOX9 . ** P
    Figure Legend Snippet: SOX9 was the target gene of miR-145. a the binding sites of miR-145 and SOX9 . b results of SOX9 enrichment, SOX9 was the target gene of miR-145. c Transfection efficiency test after the transfection of si- SOX9 or pcDNA3.1- SOX9 . ** P

    Techniques Used: Binding Assay, Transfection

    2) Product Images from "The effects of lncRNA MALAT1 on proliferation, invasion and migration in colorectal cancer through regulating SOX9"

    Article Title: The effects of lncRNA MALAT1 on proliferation, invasion and migration in colorectal cancer through regulating SOX9

    Journal: Molecular Medicine

    doi: 10.1186/s10020-018-0050-5

    Down-regulation of SOX9 inhibited the growth of tumor in vivo a the tumor specimen figure of nude mice. b statistics of tumor volume, the growth of si-MALAT1 group and si- SOX9 group was the slowest, and the volume was the smallest. The tumor volume of NC group and si- SOX9 + miR-145 inhibitor group, si-MALAT1 + pCDNA3- SOX9 group was comparative. pcDNA3.1- SOX9 group had the largest tumor volume. c Statistical results showed that tumor weight in nude mice, the tumor weight was significantly reduced in si-MALAT1 group. d The relative miR-145 expression was conspicuously increased in si-MALAT1 group in tumor tissues. e , f The qRT-PCR and western blot showed SOX9 expression was significantly decreased in si-MALAT1 group in tumor tissues. ** P
    Figure Legend Snippet: Down-regulation of SOX9 inhibited the growth of tumor in vivo a the tumor specimen figure of nude mice. b statistics of tumor volume, the growth of si-MALAT1 group and si- SOX9 group was the slowest, and the volume was the smallest. The tumor volume of NC group and si- SOX9 + miR-145 inhibitor group, si-MALAT1 + pCDNA3- SOX9 group was comparative. pcDNA3.1- SOX9 group had the largest tumor volume. c Statistical results showed that tumor weight in nude mice, the tumor weight was significantly reduced in si-MALAT1 group. d The relative miR-145 expression was conspicuously increased in si-MALAT1 group in tumor tissues. e , f The qRT-PCR and western blot showed SOX9 expression was significantly decreased in si-MALAT1 group in tumor tissues. ** P

    Techniques Used: In Vivo, Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot

    Effects of SOX9 on cycle and apoptosis of colorectal cancer cells a the cell cycle was assessed by flow cytometry, the number of cells in G1 phase was the most in the si- SOX9 group and was the least in pcDNA3.1- SOX9 group, the si- SOX9 + miR-145 inhibitor group was comparative to the NC group. b The apoptosis rate which measured by flow cytometry of pcDNA3.1- SOX9 group was the highest, si- SOX9 + miR-145 inhibitor group and NC group took the second place, and si- SOX9 group had the lowest apoptosis rate. * P
    Figure Legend Snippet: Effects of SOX9 on cycle and apoptosis of colorectal cancer cells a the cell cycle was assessed by flow cytometry, the number of cells in G1 phase was the most in the si- SOX9 group and was the least in pcDNA3.1- SOX9 group, the si- SOX9 + miR-145 inhibitor group was comparative to the NC group. b The apoptosis rate which measured by flow cytometry of pcDNA3.1- SOX9 group was the highest, si- SOX9 + miR-145 inhibitor group and NC group took the second place, and si- SOX9 group had the lowest apoptosis rate. * P

    Techniques Used: Flow Cytometry, Cytometry

    Effects of SOX9 on proliferation, invasion and migration of colorectal cancer cells a and b the cell proliferation of colorectal cancer cell lines DLD-1 and HT-29 transfected with different interference sequences, cell proliferation was the weakest in si- SOX9 group, and pcDNA3.1- SOX9 group had the strongest cell proliferation ability compared with NC group. The cell proliferation of si- SOX9 + miR-145 inhibitor group and NC group was comparative. c cell colony formation of colorectal cancer, the formation rate of colorectal cancer cells in the si- SOX9 group was remarkably slower than that of the NC group. The cell formation rate was significantly higher in pcDNA3.1- SOX9 group than that of NC group, and the cell formation in si- SOX9 + miR-145 inhibitor group and NC group had no significant difference. d invasion assays of two kinds of cancer cells under different interference conditions. e the scratch wound healing assays of two kinds of cancer cells under different interference conditions. ** P
    Figure Legend Snippet: Effects of SOX9 on proliferation, invasion and migration of colorectal cancer cells a and b the cell proliferation of colorectal cancer cell lines DLD-1 and HT-29 transfected with different interference sequences, cell proliferation was the weakest in si- SOX9 group, and pcDNA3.1- SOX9 group had the strongest cell proliferation ability compared with NC group. The cell proliferation of si- SOX9 + miR-145 inhibitor group and NC group was comparative. c cell colony formation of colorectal cancer, the formation rate of colorectal cancer cells in the si- SOX9 group was remarkably slower than that of the NC group. The cell formation rate was significantly higher in pcDNA3.1- SOX9 group than that of NC group, and the cell formation in si- SOX9 + miR-145 inhibitor group and NC group had no significant difference. d invasion assays of two kinds of cancer cells under different interference conditions. e the scratch wound healing assays of two kinds of cancer cells under different interference conditions. ** P

    Techniques Used: Migration, Transfection

    SOX9 was the target gene of miR-145. a the binding sites of miR-145 and SOX9 . b results of SOX9 enrichment, SOX9 was the target gene of miR-145. c Transfection efficiency test after the transfection of si- SOX9 or pcDNA3.1- SOX9 . ** P
    Figure Legend Snippet: SOX9 was the target gene of miR-145. a the binding sites of miR-145 and SOX9 . b results of SOX9 enrichment, SOX9 was the target gene of miR-145. c Transfection efficiency test after the transfection of si- SOX9 or pcDNA3.1- SOX9 . ** P

    Techniques Used: Binding Assay, Transfection

    MALAT1 and SOX9 were highly expressed in colorectal cancer tissues while miR-145 was low expressed a The relative MALAT1 expression in colorectal cancer tissues and adjacent tissues. b the expression of miR-145 in colorectal cancer tissues and adjacent tissues. c qRT-PCR detected the expression of SOX9 in colorectal cancer tissues and adjacent tissues. d western blot detected the expression of SOX9 in colorectal cancer tissues and adjacent tissues. ** P
    Figure Legend Snippet: MALAT1 and SOX9 were highly expressed in colorectal cancer tissues while miR-145 was low expressed a The relative MALAT1 expression in colorectal cancer tissues and adjacent tissues. b the expression of miR-145 in colorectal cancer tissues and adjacent tissues. c qRT-PCR detected the expression of SOX9 in colorectal cancer tissues and adjacent tissues. d western blot detected the expression of SOX9 in colorectal cancer tissues and adjacent tissues. ** P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    Related Articles

    Incubation:

    Article Title: Mesenchymal stem cell-derived exosomes protect trabecular meshwork from oxidative stress
    Article Snippet: .. After washing, blots were incubated with peroxidase-conjugated anti-rabbit IgG horseradish and peroxidase secondary antibodies (ab205718, 1:2000 dilution, Abcam, USA) at room temperature for 1 h. The Alpha FluorChem E system (ProteinSimple, USA) was used to visualize the protein bands. ..

    Article Title: Role of autophagy in sepsis-induced skeletal muscle dysfunction, whole-body metabolism, and survival
    Article Snippet: .. Membranes were blocked in PBS + 1% Tween® 20 + 5% BSA for 1 h at room temperature then incubated overnight with specific primary antibodies at 4°C.Membranes were washed in PBST (3×5 min) then incubated with HRP-conjugated goat anti-rabbit (ab6721) or rabbit anti-mouse (ab6728) secondary antibodies (Abcam, Cambridge, MA) for 1 h at room temperature, before further washing. ..

    Microarray:

    Article Title: VMP1, a novel prognostic biomarker, contributes to glioma development by regulating autophagy
    Article Snippet: .. Briefly, the tissue microarray slides were analyzed via immunohistochemistry with anti-human VMP1 (1:200; cat #12978S, Cell Signaling Technology, Danvers, MA, USA), followed by a horseradish peroxidase (HRP) secondary antibody (cat #ab205718; Abcam, Cambridge, UK) and DAB. ..

    Immunohistochemistry:

    Article Title: VMP1, a novel prognostic biomarker, contributes to glioma development by regulating autophagy
    Article Snippet: .. Briefly, the tissue microarray slides were analyzed via immunohistochemistry with anti-human VMP1 (1:200; cat #12978S, Cell Signaling Technology, Danvers, MA, USA), followed by a horseradish peroxidase (HRP) secondary antibody (cat #ab205718; Abcam, Cambridge, UK) and DAB. ..

    Western Blot:

    Article Title: Collagen XVII inhibits breast cancer cell proliferation and growth through deactivation of the AKT/mTOR signaling pathway
    Article Snippet: .. For western blotting, anti-collagen XVII (ab184996) and HRP-conjugated secondary antibody (ab205718) were purchased from Abcam, MA, USA. ..

    Luciferase:

    Article Title: Calcium-Sensing Receptors Control CYP27B1-Luciferase Expression: Transcriptional and Posttranscriptional Mechanisms
    Article Snippet: .. The following antibodies were used: Anti-firefly luciferase (rabbit polyclonal; Sigma #L0159; [ ]); Anti-GAPDH (rabbit monoclonal; Cell Signaling Technology #2118; [ ]); Anti-rabbit IgG-horseradish peroxidase [HRP] conjugated (goat polyclonal, Abcam #97051; [ ]); Anti-CaSR phosphoT888 (rabbit polyclonal, custom-generated and affinity purified; [ ]); Anti-CaSR total (mouse monoclonal, Thermo-Fisher #MA1-934; [ ]); Anti-rabbit Ig-HRP conjugated (goat polyclonal, Agilent #P0448; [ ]); Anti-mouse IgG-HRP conjugated (horse polyclonal, Cell Signaling Technology #7076; [ ]). ..

    Affinity Purification:

    Article Title: Calcium-Sensing Receptors Control CYP27B1-Luciferase Expression: Transcriptional and Posttranscriptional Mechanisms
    Article Snippet: .. The following antibodies were used: Anti-firefly luciferase (rabbit polyclonal; Sigma #L0159; [ ]); Anti-GAPDH (rabbit monoclonal; Cell Signaling Technology #2118; [ ]); Anti-rabbit IgG-horseradish peroxidase [HRP] conjugated (goat polyclonal, Abcam #97051; [ ]); Anti-CaSR phosphoT888 (rabbit polyclonal, custom-generated and affinity purified; [ ]); Anti-CaSR total (mouse monoclonal, Thermo-Fisher #MA1-934; [ ]); Anti-rabbit Ig-HRP conjugated (goat polyclonal, Agilent #P0448; [ ]); Anti-mouse IgG-HRP conjugated (horse polyclonal, Cell Signaling Technology #7076; [ ]). ..

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  • 99
    Abcam goat anti rabbit igg
    csPDI mediates axon repulsion in vitro a, Collapse assays testing purified bovine PDI in liposomes at a range of concentrations; controls, phosphate-buffered saline (PBS) and untreated liposomes; histogram shows mean +s.e.m. b, Assays testing PDI and GSNO applied individually or concomitantly. c, Assays testing PACMA31 and PACMA56 on somite extracts (SE). d, Assays testing reducing agents at the concentrations indicated when applied either alone or together with PDI+GSNO. e, Assays testing GSH and L-homocysteine on SE-induced collapse. f, Assays testing myoglobin (20μM) on SE- and PDI+GSNO-induced collapse. g, Assays testing carboxy (C)-PTIO (20μM) on PDI+GSNO-induced collapse. h, Assays testing L-NAME and its control D-NAME on SE-induced collapse; calcimycin was used as a positive control. i, S-nitrosylated protein (iodo-TMT-labelled) in somites; protein samples (48μg) from somite cell-free extracts were fractionated on NuPAGE 4-12% Bis Tris gels as described in the Methods; lanes 1 2 are controls consisting of somite proteins only, with no detectable signal compared with lanes 3 4 where GSNO has been added; lane 3 is a control (treated with water) showing negligible iodoTMT labelling, and lane 4 (reduced with ascorbate to generate a new free thiol for labelling) shows increased label; lane 5 shows that addition of PDI (1μg/0.1ml reaction mixture) enhances labelling; lane 6 shows that 3mM GSH in the absence of GSNO and PDI does not generate a signal; lane 7 shows that 3μM GSH is insufficient to interfere with nitrosylation, concurring with the findings of Sliskovic et al. 37 . The coloured molecular weight markers on the blot are shown on the left (BLUeye prestained protein ladder, 2.5 μL, Geneflow). j, Identification of LC1 in somite extract (25μg protein); the blot was cut in half above the 41K marker and the top half of the membrane was probed with rabbit <t>anti-tubulin</t> followed by goat anti-rabbit <t>IgG;</t> the bottom half was probed with mouse monoclonal antibody against amino acids 2257-2357 of mouse MAP-1B (LC1) followed by goat anti-mouse IgG. The molecular weight markers on the blot are shown to the right (BLUeye prestained protein ladder, 3μl). k, Identification of LC1 as a substrate for S-nitrosylation; cell-free somite extract (200μg) was treated with D-NAME or L-NAME, followed by further incubation in GSNO (200μM), as described in the Methods. Samples were then processed for the presence of S-nitrosylated proteins using iodo-TMT as described in the Methods. Protein samples (15μg) were then fractionated and blotted, after which the blot was cut as described for Fig. 3j . The top half was treated with anti-tubulin and the bottom half with anti-iodoTMT. L-NAME treatment blocked S-nitrosylation, as shown by the lack of iodoTMT labelling. The control D-NAME was without effect.
    Goat Anti Rabbit Igg, 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/goat anti rabbit igg/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg - by Bioz Stars, 2021-09
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    97
    Abcam mmp 16
    Identification of interaction between miR-215 and <t>MMP-16.</t> Results of the dual-luciferase reporter assay. Plasmids (0.5 µg) with WT or mutant 3′-untranslated region DNA sequences were co-transfected with miR-215 mimics into A549 cells. Following cultivation for 24 h, the cells were lysed using a dual-luciferase reporter assay kit and fluorescence intensity was measured using GloMax 20/20 luminometer. Using Renilla fluorescence activity as an internal reference, the fluorescence values of each group of cells were measured. *P
    Mmp 16, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmp 16/product/Abcam
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    csPDI mediates axon repulsion in vitro a, Collapse assays testing purified bovine PDI in liposomes at a range of concentrations; controls, phosphate-buffered saline (PBS) and untreated liposomes; histogram shows mean +s.e.m. b, Assays testing PDI and GSNO applied individually or concomitantly. c, Assays testing PACMA31 and PACMA56 on somite extracts (SE). d, Assays testing reducing agents at the concentrations indicated when applied either alone or together with PDI+GSNO. e, Assays testing GSH and L-homocysteine on SE-induced collapse. f, Assays testing myoglobin (20μM) on SE- and PDI+GSNO-induced collapse. g, Assays testing carboxy (C)-PTIO (20μM) on PDI+GSNO-induced collapse. h, Assays testing L-NAME and its control D-NAME on SE-induced collapse; calcimycin was used as a positive control. i, S-nitrosylated protein (iodo-TMT-labelled) in somites; protein samples (48μg) from somite cell-free extracts were fractionated on NuPAGE 4-12% Bis Tris gels as described in the Methods; lanes 1 2 are controls consisting of somite proteins only, with no detectable signal compared with lanes 3 4 where GSNO has been added; lane 3 is a control (treated with water) showing negligible iodoTMT labelling, and lane 4 (reduced with ascorbate to generate a new free thiol for labelling) shows increased label; lane 5 shows that addition of PDI (1μg/0.1ml reaction mixture) enhances labelling; lane 6 shows that 3mM GSH in the absence of GSNO and PDI does not generate a signal; lane 7 shows that 3μM GSH is insufficient to interfere with nitrosylation, concurring with the findings of Sliskovic et al. 37 . The coloured molecular weight markers on the blot are shown on the left (BLUeye prestained protein ladder, 2.5 μL, Geneflow). j, Identification of LC1 in somite extract (25μg protein); the blot was cut in half above the 41K marker and the top half of the membrane was probed with rabbit anti-tubulin followed by goat anti-rabbit IgG; the bottom half was probed with mouse monoclonal antibody against amino acids 2257-2357 of mouse MAP-1B (LC1) followed by goat anti-mouse IgG. The molecular weight markers on the blot are shown to the right (BLUeye prestained protein ladder, 3μl). k, Identification of LC1 as a substrate for S-nitrosylation; cell-free somite extract (200μg) was treated with D-NAME or L-NAME, followed by further incubation in GSNO (200μM), as described in the Methods. Samples were then processed for the presence of S-nitrosylated proteins using iodo-TMT as described in the Methods. Protein samples (15μg) were then fractionated and blotted, after which the blot was cut as described for Fig. 3j . The top half was treated with anti-tubulin and the bottom half with anti-iodoTMT. L-NAME treatment blocked S-nitrosylation, as shown by the lack of iodoTMT labelling. The control D-NAME was without effect.

    Journal: bioRxiv

    Article Title: Regulation of nerve growth and patterning by cell surface protein disulphide isomerase

    doi: 10.1101/838771

    Figure Lengend Snippet: csPDI mediates axon repulsion in vitro a, Collapse assays testing purified bovine PDI in liposomes at a range of concentrations; controls, phosphate-buffered saline (PBS) and untreated liposomes; histogram shows mean +s.e.m. b, Assays testing PDI and GSNO applied individually or concomitantly. c, Assays testing PACMA31 and PACMA56 on somite extracts (SE). d, Assays testing reducing agents at the concentrations indicated when applied either alone or together with PDI+GSNO. e, Assays testing GSH and L-homocysteine on SE-induced collapse. f, Assays testing myoglobin (20μM) on SE- and PDI+GSNO-induced collapse. g, Assays testing carboxy (C)-PTIO (20μM) on PDI+GSNO-induced collapse. h, Assays testing L-NAME and its control D-NAME on SE-induced collapse; calcimycin was used as a positive control. i, S-nitrosylated protein (iodo-TMT-labelled) in somites; protein samples (48μg) from somite cell-free extracts were fractionated on NuPAGE 4-12% Bis Tris gels as described in the Methods; lanes 1 2 are controls consisting of somite proteins only, with no detectable signal compared with lanes 3 4 where GSNO has been added; lane 3 is a control (treated with water) showing negligible iodoTMT labelling, and lane 4 (reduced with ascorbate to generate a new free thiol for labelling) shows increased label; lane 5 shows that addition of PDI (1μg/0.1ml reaction mixture) enhances labelling; lane 6 shows that 3mM GSH in the absence of GSNO and PDI does not generate a signal; lane 7 shows that 3μM GSH is insufficient to interfere with nitrosylation, concurring with the findings of Sliskovic et al. 37 . The coloured molecular weight markers on the blot are shown on the left (BLUeye prestained protein ladder, 2.5 μL, Geneflow). j, Identification of LC1 in somite extract (25μg protein); the blot was cut in half above the 41K marker and the top half of the membrane was probed with rabbit anti-tubulin followed by goat anti-rabbit IgG; the bottom half was probed with mouse monoclonal antibody against amino acids 2257-2357 of mouse MAP-1B (LC1) followed by goat anti-mouse IgG. The molecular weight markers on the blot are shown to the right (BLUeye prestained protein ladder, 3μl). k, Identification of LC1 as a substrate for S-nitrosylation; cell-free somite extract (200μg) was treated with D-NAME or L-NAME, followed by further incubation in GSNO (200μM), as described in the Methods. Samples were then processed for the presence of S-nitrosylated proteins using iodo-TMT as described in the Methods. Protein samples (15μg) were then fractionated and blotted, after which the blot was cut as described for Fig. 3j . The top half was treated with anti-tubulin and the bottom half with anti-iodoTMT. L-NAME treatment blocked S-nitrosylation, as shown by the lack of iodoTMT labelling. The control D-NAME was without effect.

    Article Snippet: The blot was cut in half below the 53K molecular weight marker and the top half probed with rabbit anti-tubulin (Sigma, Lot#50K4813, 1:20,000) and goat anti-rabbit IgG (HRP, Abcam Lot#GR3231028-7) followed by Millipore Immobilon Western Reagent.

    Techniques: In Vitro, Purification, Positive Control, Molecular Weight, Marker, Incubation

    Identification of interaction between miR-215 and MMP-16. Results of the dual-luciferase reporter assay. Plasmids (0.5 µg) with WT or mutant 3′-untranslated region DNA sequences were co-transfected with miR-215 mimics into A549 cells. Following cultivation for 24 h, the cells were lysed using a dual-luciferase reporter assay kit and fluorescence intensity was measured using GloMax 20/20 luminometer. Using Renilla fluorescence activity as an internal reference, the fluorescence values of each group of cells were measured. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: MicroRNA-215 suppresses the proliferation, migration and invasion of non-small cell lung carcinoma cells through the downregulation of matrix metalloproteinase-16 expression

    doi: 10.3892/etm.2018.5869

    Figure Lengend Snippet: Identification of interaction between miR-215 and MMP-16. Results of the dual-luciferase reporter assay. Plasmids (0.5 µg) with WT or mutant 3′-untranslated region DNA sequences were co-transfected with miR-215 mimics into A549 cells. Following cultivation for 24 h, the cells were lysed using a dual-luciferase reporter assay kit and fluorescence intensity was measured using GloMax 20/20 luminometer. Using Renilla fluorescence activity as an internal reference, the fluorescence values of each group of cells were measured. *P

    Article Snippet: Following extensive washing with PBS with Tween-20 (five times, 5 min each), the membranes were incubated, respectively, with goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody for GAPDH (1:5,000; cat. no. ab6789; Abcam) and goat anti-rabbit HRP-conjugated secondary antibody for MMP-16 (1:2,000; cat. no. ab205718; Abcam) for 1 h at room temperature.

    Techniques: Luciferase, Reporter Assay, Mutagenesis, Transfection, Fluorescence, Activity Assay

    Effect of MMP-16 on the biological functions of A549 cells. (A) Expression of MMP-16 protein in A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. Western blotting was used to determine protein expression. (B) Proliferation of A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. A Cell Counting Kit-8 assay was used to measure the absorbance (490 nm) of the cells at 0, 24 and 48 h. A cell proliferation curve was plotted. (C) Migration and invasion of A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. Migratory and invasive abilities of the cells were determined using Transwell assays. Magnification, ×100. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: MicroRNA-215 suppresses the proliferation, migration and invasion of non-small cell lung carcinoma cells through the downregulation of matrix metalloproteinase-16 expression

    doi: 10.3892/etm.2018.5869

    Figure Lengend Snippet: Effect of MMP-16 on the biological functions of A549 cells. (A) Expression of MMP-16 protein in A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. Western blotting was used to determine protein expression. (B) Proliferation of A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. A Cell Counting Kit-8 assay was used to measure the absorbance (490 nm) of the cells at 0, 24 and 48 h. A cell proliferation curve was plotted. (C) Migration and invasion of A549 cells transfected with NC or pcDNA3.1-MMP-16 plasmids. Migratory and invasive abilities of the cells were determined using Transwell assays. Magnification, ×100. *P

    Article Snippet: Following extensive washing with PBS with Tween-20 (five times, 5 min each), the membranes were incubated, respectively, with goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody for GAPDH (1:5,000; cat. no. ab6789; Abcam) and goat anti-rabbit HRP-conjugated secondary antibody for MMP-16 (1:2,000; cat. no. ab205718; Abcam) for 1 h at room temperature.

    Techniques: Expressing, Transfection, Western Blot, Cell Counting, Migration

    Effect of miR-215 and MMP-16 on the biological functions of A549 cells. (A) Expression of MMP-16 protein in A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. Western blotting was used to measure protein expression. (B) Proliferation of A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. A Cell Counting Kit-8 assay was used to measure the absorbance (490 nm) of the cells at 0, 24 and 48 h. A cell proliferation curve was plotted. (C) Migration and invasion of A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. Migratory and invasive abilities of the cells were determined using Transwell assays. Magnification, ×100. The data in multiple groups for migration or invasion assays were compared using one way analysis of variance followed by a post hoc test. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: MicroRNA-215 suppresses the proliferation, migration and invasion of non-small cell lung carcinoma cells through the downregulation of matrix metalloproteinase-16 expression

    doi: 10.3892/etm.2018.5869

    Figure Lengend Snippet: Effect of miR-215 and MMP-16 on the biological functions of A549 cells. (A) Expression of MMP-16 protein in A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. Western blotting was used to measure protein expression. (B) Proliferation of A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. A Cell Counting Kit-8 assay was used to measure the absorbance (490 nm) of the cells at 0, 24 and 48 h. A cell proliferation curve was plotted. (C) Migration and invasion of A549 cells transfected with NC, miR-215 or miR-215 and pcDNA3.1-MMP-16 plasmids. Migratory and invasive abilities of the cells were determined using Transwell assays. Magnification, ×100. The data in multiple groups for migration or invasion assays were compared using one way analysis of variance followed by a post hoc test. *P

    Article Snippet: Following extensive washing with PBS with Tween-20 (five times, 5 min each), the membranes were incubated, respectively, with goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody for GAPDH (1:5,000; cat. no. ab6789; Abcam) and goat anti-rabbit HRP-conjugated secondary antibody for MMP-16 (1:2,000; cat. no. ab205718; Abcam) for 1 h at room temperature.

    Techniques: Expressing, Transfection, Western Blot, Cell Counting, Migration

    Effect of miR-215 on the expression of MMP-16 protein in A549 cells. Western blotting was used to determine protein expression. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: MicroRNA-215 suppresses the proliferation, migration and invasion of non-small cell lung carcinoma cells through the downregulation of matrix metalloproteinase-16 expression

    doi: 10.3892/etm.2018.5869

    Figure Lengend Snippet: Effect of miR-215 on the expression of MMP-16 protein in A549 cells. Western blotting was used to determine protein expression. *P

    Article Snippet: Following extensive washing with PBS with Tween-20 (five times, 5 min each), the membranes were incubated, respectively, with goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody for GAPDH (1:5,000; cat. no. ab6789; Abcam) and goat anti-rabbit HRP-conjugated secondary antibody for MMP-16 (1:2,000; cat. no. ab205718; Abcam) for 1 h at room temperature.

    Techniques: Expressing, Western Blot

    Ablation of HTLV-1 CTCF-binding site significantly decreases HTLV-1-specific antibody response, but not total rabbit IgG. a Antibody response was quantified using a modified Avioq HTLV-1/2 Microelisa System protocol (Avioq, Inc., Research Triangle Park, NC). The supplied horseradish peroxidase (HRP) conjugated goat anti-human immunoglobulin (Ig) was substituted for an HRP conjugated goat anti-rabbit IgG (ab6721; Abcam, Cambridge, United Kingdom). Rabbit plasma was diluted 1:500 to obtain absorbance values within the linear range of the assay. Each symbol represents the absorbance value of a single inoculated rabbit at 0, 2, 4, 8, or 12 weeks post-infection within each group. b Total rabbit IgG was quantified using the Abcam Rabbit IgG ELISA Kit in accordance with the provided protocol (ab187400; Abcam, Cambridge, United Kingdom). Plasma samples were diluted 1:1 × 10 6 . Each symbol represents total IgG of a single inoculated rabbit at 0, 2, or 12 weeks post-infection within each group. Bars represent mean absorbance or IgG values. Mixed model analyses with a Bonferroni correction were performed in weeks 8 and 12 (HTLV-1-specific) or 2 and 12 (total rabbit IgG) to determine statistical significance. A p

    Journal: Retrovirology

    Article Title: HTLV-1 CTCF-binding site is dispensable for in vitro immortalization and persistent infection in vivo

    doi: 10.1186/s12977-019-0507-9

    Figure Lengend Snippet: Ablation of HTLV-1 CTCF-binding site significantly decreases HTLV-1-specific antibody response, but not total rabbit IgG. a Antibody response was quantified using a modified Avioq HTLV-1/2 Microelisa System protocol (Avioq, Inc., Research Triangle Park, NC). The supplied horseradish peroxidase (HRP) conjugated goat anti-human immunoglobulin (Ig) was substituted for an HRP conjugated goat anti-rabbit IgG (ab6721; Abcam, Cambridge, United Kingdom). Rabbit plasma was diluted 1:500 to obtain absorbance values within the linear range of the assay. Each symbol represents the absorbance value of a single inoculated rabbit at 0, 2, 4, 8, or 12 weeks post-infection within each group. b Total rabbit IgG was quantified using the Abcam Rabbit IgG ELISA Kit in accordance with the provided protocol (ab187400; Abcam, Cambridge, United Kingdom). Plasma samples were diluted 1:1 × 10 6 . Each symbol represents total IgG of a single inoculated rabbit at 0, 2, or 12 weeks post-infection within each group. Bars represent mean absorbance or IgG values. Mixed model analyses with a Bonferroni correction were performed in weeks 8 and 12 (HTLV-1-specific) or 2 and 12 (total rabbit IgG) to determine statistical significance. A p

    Article Snippet: The supplied horseradish peroxidase (HRP) conjugated goat anti-human IgG was substituted for an HRP conjugated goat anti-rabbit IgG (ab6721; Abcam, Cambridge, United Kingdom).

    Techniques: Binding Assay, Modification, Infection, Enzyme-linked Immunosorbent Assay

    Circ_DOCK1 expression is enhanced in colorectal cancer samples and cell lines. a Circ_DOCK1 expression was determined using qRT-PCR in tumor and normal samples. n =42. b Circ_DOCK1 level was measured via qRT-PCR in HCT116, SW480, and FHC cells. c , d Circ_DOCK1 and DOCK1 levels were examined via qRT-PCR in HCT116 and SW480 cells after actinomycin D treatment for 0, 8, 16, or 24 h. e , f Circ_DOCK1, β-actin, and U6 levels were measured via qRT-PCR in nucleus and cytoplasm of HCT116 and SW480 cells. * P

    Journal: World Journal of Surgical Oncology

    Article Title: Circ_DOCK1 regulates USP11 through miR-132-3p to control colorectal cancer progression

    doi: 10.1186/s12957-021-02173-x

    Figure Lengend Snippet: Circ_DOCK1 expression is enhanced in colorectal cancer samples and cell lines. a Circ_DOCK1 expression was determined using qRT-PCR in tumor and normal samples. n =42. b Circ_DOCK1 level was measured via qRT-PCR in HCT116, SW480, and FHC cells. c , d Circ_DOCK1 and DOCK1 levels were examined via qRT-PCR in HCT116 and SW480 cells after actinomycin D treatment for 0, 8, 16, or 24 h. e , f Circ_DOCK1, β-actin, and U6 levels were measured via qRT-PCR in nucleus and cytoplasm of HCT116 and SW480 cells. * P

    Article Snippet: The membranes were blocked in the blocking buffer (Beyotime) for 1 h, and then incubated with primary antibodies for anti-CyclinD1 (ab226977, 1:300 dilution, Abcam, Cambridge, UK), anti-USP11 (ab109232, 1:3000 dilution, Abcam), or anti-β-actin (ab8227, 1:3000 dilution, Abcam) overnight, followed by incubation of horseradish peroxidase (HRP)-labeled IgG (ab6721, 1:10000 dilution, Abcam) for 2 h. β-actin was used as a loading control.

    Techniques: Expressing, Quantitative RT-PCR