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  • 86
    Thermo Fisher gene exp spc24 hs00699347 m1
    Gene Exp Spc24 Hs00699347 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    spc24  (Abcam)
    91
    Abcam spc24
    <t>SPC24</t> over-expression is observed in lung tumors of whom the patients are smokers Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from smokers vs. non-smokers. Four Oncomine expression analyses are shown.
    Spc24, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam anti spc24 rabbit ab
    <t>SPC24</t> protein was analyzed by immunohistochemistry and western blot in HCC tissues A and B. The representative immunohistochemical pictures of one pair of HCC specimen (upper panel) and corresponding noncancerous tissue (lower panel) from a tissue array containing 69 pairs of HCC specimens were shown. The pictures were stainied with SPC24 antibody, and the nuclei were counterstained with hematoxylin. C. The representative immunohistochemical staining in one normal liver tissue is shown. The nuclei were counterstained with hematoxylin. D. SPC24 expression in 2 normal liver tissues, 2 paired HCC [C] and adjacent non-cancerous liver tissues [N] was evaluated by Western blot. β-actin protein was used as an internal control.
    Anti Spc24 Rabbit Ab, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti spc24 rabbit ab/product/Abcam
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    90
    GenePharma Company spc24
    Knockdown of <t>Spc24</t> causes aneuploidy ( A ) Oocytes in MII stage were collected and chromosome spreads were performed in control and Spc24 RNAi oocytes. Scale bars: 20 μm. ( B ) The numbers of univalents in the oocytes in A a; b; c are 20; 19; 23, (“a” is control oocyte, “b,c” are RNAi oocytes), respectively. The total numbers of analyzed oocytes are indicated (n).
    Spc24, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/spc24/product/GenePharma Company
    Average 90 stars, based on 5 article reviews
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    91
    OriGene spc24
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    Spc24, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/spc24/product/OriGene
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    91
    Shanghai GenePharma spc24 spc24 small interference rnas sirnas
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    Spc24 Spc24 Small Interference Rnas Sirnas, supplied by Shanghai GenePharma, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology polyclonal anti spc24 antibody
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    Polyclonal Anti Spc24 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bethyl rabbit anti spc24 antibody affinity purified
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    Rabbit Anti Spc24 Antibody Affinity Purified, supplied by Bethyl, used in various techniques. Bioz Stars score: 91/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore b mori 6xhis spc24 73 162 spc25 70 211
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    B Mori 6xhis Spc24 73 162 Spc25 70 211, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore prare sigma 71400 b mori spc24 spc25 baculovirus
    <t>SPC24</t> knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p
    Prare Sigma 71400 B Mori Spc24 Spc25 Baculovirus, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SPC24 over-expression is observed in lung tumors of whom the patients are smokers Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from smokers vs. non-smokers. Four Oncomine expression analyses are shown.

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: SPC24 over-expression is observed in lung tumors of whom the patients are smokers Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from smokers vs. non-smokers. Four Oncomine expression analyses are shown.

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Over Expression, Expressing

    SPC24 over-expression is associated with lung tumors from patients with short survival Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from patients who died at 1, 3, or 5 years after diagnosis, compared with those who were still alive at the same time point.

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: SPC24 over-expression is associated with lung tumors from patients with short survival Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from patients who died at 1, 3, or 5 years after diagnosis, compared with those who were still alive at the same time point.

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Over Expression

    Knocking down SPC24 represses cell growth and promotes apoptosis in lung cancer cell lines ( A ) SPC24 was knocked down by siRNA in PC9 and A549 cells. Western blot of SPC24 was analyzed in the knockdown (si SPC24 ) and control (siN) cells. ( B ) Cell growth for SPC24 -knockdown and control cells was measured as viable cell numbers that was recorded as OD 450 at 0, 24, 72, and 96 h. ( C ) Apoptosis was recorded for knockdown and control cells by flow cytometry analysis of Annexin V-FITC-Propidium Iodide (PI) stained populations.

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: Knocking down SPC24 represses cell growth and promotes apoptosis in lung cancer cell lines ( A ) SPC24 was knocked down by siRNA in PC9 and A549 cells. Western blot of SPC24 was analyzed in the knockdown (si SPC24 ) and control (siN) cells. ( B ) Cell growth for SPC24 -knockdown and control cells was measured as viable cell numbers that was recorded as OD 450 at 0, 24, 72, and 96 h. ( C ) Apoptosis was recorded for knockdown and control cells by flow cytometry analysis of Annexin V-FITC-Propidium Iodide (PI) stained populations.

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Western Blot, Flow Cytometry, Cytometry, Staining

    Knocking down SPC24 suppresses cellular migration in lung cancer cell lines ( A ) Western blots of the EMT markers, including E-cadherin, Vimentin, N-cadherin,along with SPC24, are shown in SPC24-knockdown and control cells. ( B ) Migration of the knockdown and control cells was evaluated by Transwell migration assay. ( C ) Quantification of the cellular migration shown in (B). (Mean ± S.D., n = 3).

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: Knocking down SPC24 suppresses cellular migration in lung cancer cell lines ( A ) Western blots of the EMT markers, including E-cadherin, Vimentin, N-cadherin,along with SPC24, are shown in SPC24-knockdown and control cells. ( B ) Migration of the knockdown and control cells was evaluated by Transwell migration assay. ( C ) Quantification of the cellular migration shown in (B). (Mean ± S.D., n = 3).

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Migration, Western Blot, Transwell Migration Assay

    Knocking down SPC24 inhibits tumor growth in vivo ( A ) Stable SPC24 - knockdown (sh SPC24 ) cell line of PC9 was generated by shRNA, and injected subcutaneously into 6-week old immunocompromised nude mice. At time of sacrifice, tumors were collected and weighed. ( B ) The weights of tumors are recorded as Mean ± S.D. ( n = 3) * p

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: Knocking down SPC24 inhibits tumor growth in vivo ( A ) Stable SPC24 - knockdown (sh SPC24 ) cell line of PC9 was generated by shRNA, and injected subcutaneously into 6-week old immunocompromised nude mice. At time of sacrifice, tumors were collected and weighed. ( B ) The weights of tumors are recorded as Mean ± S.D. ( n = 3) * p

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: In Vivo, Generated, shRNA, Injection, Mouse Assay

    SPC24 over-expression is associated with lung tumors from patients with recurrence Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from patients with or without recurrent cancer. Four Oncomine expression analyses are shown.

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: SPC24 over-expression is associated with lung tumors from patients with recurrence Oncomine boxed plots of SPC24 levels in lung adenocarcinomas from patients with or without recurrent cancer. Four Oncomine expression analyses are shown.

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Over Expression, Expressing

    SPC24 is over-expressed in human lung adenocarcinoma tumors ( A ) Oncomine boxed plots of SPC24 in human adenocarcinoma and normal lungs. Oncomine data resources are listed under each plot. ( B ) IHC analysis of SPC24 in clinical normal and lung cancer samples, representing weak staining in the normal tissues, and strong one in the tumors. Ten ( N = 10) tumors and 8 normal samples were examined. The staining signals are classified as weak, medium, and strong, and tabulated as the statistics for normal and tumor tissues. The χ 2 statistic is 128.808, with p-value

    Journal: Oncotarget

    Article Title: A potential prognostic biomarker SPC24 promotes tumorigenesis and metastasis in lung cancer

    doi: 10.18632/oncotarget.18971

    Figure Lengend Snippet: SPC24 is over-expressed in human lung adenocarcinoma tumors ( A ) Oncomine boxed plots of SPC24 in human adenocarcinoma and normal lungs. Oncomine data resources are listed under each plot. ( B ) IHC analysis of SPC24 in clinical normal and lung cancer samples, representing weak staining in the normal tissues, and strong one in the tumors. Ten ( N = 10) tumors and 8 normal samples were examined. The staining signals are classified as weak, medium, and strong, and tabulated as the statistics for normal and tumor tissues. The χ 2 statistic is 128.808, with p-value

    Article Snippet: SPC24 (Cat. ab169786), E-cadherin (Cat. ab1416), and β-actin (Cat. ab8226) antibodies were purchased from Abcam.

    Techniques: Immunohistochemistry, Staining

    SPC24 protein was analyzed by immunohistochemistry and western blot in HCC tissues A and B. The representative immunohistochemical pictures of one pair of HCC specimen (upper panel) and corresponding noncancerous tissue (lower panel) from a tissue array containing 69 pairs of HCC specimens were shown. The pictures were stainied with SPC24 antibody, and the nuclei were counterstained with hematoxylin. C. The representative immunohistochemical staining in one normal liver tissue is shown. The nuclei were counterstained with hematoxylin. D. SPC24 expression in 2 normal liver tissues, 2 paired HCC [C] and adjacent non-cancerous liver tissues [N] was evaluated by Western blot. β-actin protein was used as an internal control.

    Journal: Oncotarget

    Article Title: A novel prognostic biomarker SPC24 up-regulated in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: SPC24 protein was analyzed by immunohistochemistry and western blot in HCC tissues A and B. The representative immunohistochemical pictures of one pair of HCC specimen (upper panel) and corresponding noncancerous tissue (lower panel) from a tissue array containing 69 pairs of HCC specimens were shown. The pictures were stainied with SPC24 antibody, and the nuclei were counterstained with hematoxylin. C. The representative immunohistochemical staining in one normal liver tissue is shown. The nuclei were counterstained with hematoxylin. D. SPC24 expression in 2 normal liver tissues, 2 paired HCC [C] and adjacent non-cancerous liver tissues [N] was evaluated by Western blot. β-actin protein was used as an internal control.

    Article Snippet: Sections were incubated with an anti-SPC24 rabbit Ab (1:200 dilution, lot GR147853–1, Abcam) overnight in a humidified container at 4°C, the next day, after being washed with PBS, a secondary goat anti-rabbit antibody conjugated to horseradish peroxidase was applied for 1 hour at room temperature.

    Techniques: Immunohistochemistry, Western Blot, Staining, Expressing

    SPC24 inhibition affected HCC cell adhesion, invasion, and apoptosis SPC24 inhibition by siRNA-2 dramatically reduced cell adhesive ratio in both SMMC-7721 and HepG2 cells A. The invasiveness was evaluated by Matrigel assay in which scrambled siRNA served as a control and the cell number represented the mean values per field (from at least five fields) from 3 independent experiments (right panel) (mean ± SD), the data showed that the invasion of SMMC-7721 B. and HepG2 C. cells transfected with siRNA-2 was dramatically inhibited. Additionally, cleave d caspase-3 (active form of caspase-3) in SMMC7721 and HepG2 cells was increased by transfection with siRNA-2 confirmed by Western blot D.

    Journal: Oncotarget

    Article Title: A novel prognostic biomarker SPC24 up-regulated in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: SPC24 inhibition affected HCC cell adhesion, invasion, and apoptosis SPC24 inhibition by siRNA-2 dramatically reduced cell adhesive ratio in both SMMC-7721 and HepG2 cells A. The invasiveness was evaluated by Matrigel assay in which scrambled siRNA served as a control and the cell number represented the mean values per field (from at least five fields) from 3 independent experiments (right panel) (mean ± SD), the data showed that the invasion of SMMC-7721 B. and HepG2 C. cells transfected with siRNA-2 was dramatically inhibited. Additionally, cleave d caspase-3 (active form of caspase-3) in SMMC7721 and HepG2 cells was increased by transfection with siRNA-2 confirmed by Western blot D.

    Article Snippet: Sections were incubated with an anti-SPC24 rabbit Ab (1:200 dilution, lot GR147853–1, Abcam) overnight in a humidified container at 4°C, the next day, after being washed with PBS, a secondary goat anti-rabbit antibody conjugated to horseradish peroxidase was applied for 1 hour at room temperature.

    Techniques: Inhibition, Matrigel Assay, Transfection, Western Blot

    SPC24 mRNA expression in HCC specimens analyzed by RT-PCR and real-time RT-PCR A. The mRNA of SPC24 was measured in 20 cases of HCC specimens [C] and corresponding adjacent noncancerous liver tissues [N] by RT-PCR. The relative mRNA level of SPC24 was normalized based on that of an internal reference β-actin. PCR products were visualized by electrophoresis on 2% agarose gels. B. The mRNA of SPC24 was evaluated in 9 cases of normal liver tissues by semi-quantitative RT-PCR. β-Actin was used as an internal control. PCR products were visualized after electrophoresis through 2% agarose. C. The mRNA levels of SPC24 in 212 paired HCC specimens and adjacent noncancerous liver tissues (ANLT) were determined by quantitative real-time PCR, β-Actin was used as an internal control. D. Combined analysis of SPC24 mRNA expression and AFP level in examined 212 cases of HCC. The number represents the percentage among the 212 cases of HCC patients.

    Journal: Oncotarget

    Article Title: A novel prognostic biomarker SPC24 up-regulated in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: SPC24 mRNA expression in HCC specimens analyzed by RT-PCR and real-time RT-PCR A. The mRNA of SPC24 was measured in 20 cases of HCC specimens [C] and corresponding adjacent noncancerous liver tissues [N] by RT-PCR. The relative mRNA level of SPC24 was normalized based on that of an internal reference β-actin. PCR products were visualized by electrophoresis on 2% agarose gels. B. The mRNA of SPC24 was evaluated in 9 cases of normal liver tissues by semi-quantitative RT-PCR. β-Actin was used as an internal control. PCR products were visualized after electrophoresis through 2% agarose. C. The mRNA levels of SPC24 in 212 paired HCC specimens and adjacent noncancerous liver tissues (ANLT) were determined by quantitative real-time PCR, β-Actin was used as an internal control. D. Combined analysis of SPC24 mRNA expression and AFP level in examined 212 cases of HCC. The number represents the percentage among the 212 cases of HCC patients.

    Article Snippet: Sections were incubated with an anti-SPC24 rabbit Ab (1:200 dilution, lot GR147853–1, Abcam) overnight in a humidified container at 4°C, the next day, after being washed with PBS, a secondary goat anti-rabbit antibody conjugated to horseradish peroxidase was applied for 1 hour at room temperature.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Polymerase Chain Reaction, Electrophoresis, Real-time Polymerase Chain Reaction

    SPC expression affected disease-free survival rate and overall survival rate in patients with HCC All patients were divided into two groups according to the expression level of SPC24, then the prognostic significance of SPC24 was evaluated by Kaplan-Meier survival analysis and log-rank test. A. DFS of patients with high expression of SPC24 was shorter than that with low expression of SPC24. B. OS of patients with high expression of SPC24 was shorter than that with low expression of SPC24. C and D. High expression of SPC24 in HCC with diameter of tumor size > 5 cm appeared apparent prognostic value in predicting poorer DFS and OS. E and F. High expression of SPC24 in HCC patients with one single tumor nodule was significantly correlated with shorter DFS and OS.

    Journal: Oncotarget

    Article Title: A novel prognostic biomarker SPC24 up-regulated in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: SPC expression affected disease-free survival rate and overall survival rate in patients with HCC All patients were divided into two groups according to the expression level of SPC24, then the prognostic significance of SPC24 was evaluated by Kaplan-Meier survival analysis and log-rank test. A. DFS of patients with high expression of SPC24 was shorter than that with low expression of SPC24. B. OS of patients with high expression of SPC24 was shorter than that with low expression of SPC24. C and D. High expression of SPC24 in HCC with diameter of tumor size > 5 cm appeared apparent prognostic value in predicting poorer DFS and OS. E and F. High expression of SPC24 in HCC patients with one single tumor nodule was significantly correlated with shorter DFS and OS.

    Article Snippet: Sections were incubated with an anti-SPC24 rabbit Ab (1:200 dilution, lot GR147853–1, Abcam) overnight in a humidified container at 4°C, the next day, after being washed with PBS, a secondary goat anti-rabbit antibody conjugated to horseradish peroxidase was applied for 1 hour at room temperature.

    Techniques: Expressing

    The inhibition of SPC24 decreased HCC cell growth SPC24 mRNA was detected by semi-quantitative RT-PCR; the products were visualized after electrophoresis on 2% agarose gels and β-actin served as an interval control. mRNA levels of SPC24 were detected by semi-quantitative RT-PCR in 12 kinds of human HCC cell lines and one of normal liver cell line A. RNAi efficiency was demonstrated by semi-quantitative RT-PCR in SMMC-7721 B. and HepG2 C. cells, and also confirmed by Western blot in SMMC-7721 D. and HepG2 E. cells. SPC24 inhibition by siRNA-2 dramatically decreased cell growth in both SMMC-7721 F. and HepG2 G. cells using cell viability assay. Experiments were performed in triplicate and data were shown as mean ± SD, * p

    Journal: Oncotarget

    Article Title: A novel prognostic biomarker SPC24 up-regulated in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: The inhibition of SPC24 decreased HCC cell growth SPC24 mRNA was detected by semi-quantitative RT-PCR; the products were visualized after electrophoresis on 2% agarose gels and β-actin served as an interval control. mRNA levels of SPC24 were detected by semi-quantitative RT-PCR in 12 kinds of human HCC cell lines and one of normal liver cell line A. RNAi efficiency was demonstrated by semi-quantitative RT-PCR in SMMC-7721 B. and HepG2 C. cells, and also confirmed by Western blot in SMMC-7721 D. and HepG2 E. cells. SPC24 inhibition by siRNA-2 dramatically decreased cell growth in both SMMC-7721 F. and HepG2 G. cells using cell viability assay. Experiments were performed in triplicate and data were shown as mean ± SD, * p

    Article Snippet: Sections were incubated with an anti-SPC24 rabbit Ab (1:200 dilution, lot GR147853–1, Abcam) overnight in a humidified container at 4°C, the next day, after being washed with PBS, a secondary goat anti-rabbit antibody conjugated to horseradish peroxidase was applied for 1 hour at room temperature.

    Techniques: Inhibition, Quantitative RT-PCR, Electrophoresis, Western Blot, Viability Assay

    The Ndc80 complex interacts with Bod1 in mitotic HeLa cell extracts and in vitro . ( a ) Graphic representation of the full-length Ndc80 complex. Domains of interest, including the microtubule and centromere binding regions of the complex, are labelled. ( b ) Co-precipitation of Ndc80 complex components Ndc80, Nuf2 and Spc24 from mitotic HeLa cell extracts with purified Bod1-GST. ( c ) Representation of the recombinant Ndc80 Bonsai complex (as described in [ 38 ]). Ndc80–Spc25 and GST–Nuf2–Spc24 are expressed as fusion proteins. Both fusion constructs are co-expressed in E. coli from a dual-expression vector. ( d ) Of note, 150 pmol recombinant Ndc80 Bonsai, consisting of dimers of one Ndc80–Spc25 fusion protein with one GST–Nuf2–Spc24 fusion protein, was immobilized on Sepharose beads and incubated with 1 nmol Bod1-MBP or MBP. Binding was allowed for 1 h. Proteins were resolved by SDS-PAGE and immunoblotted using simultaneous detection of the MBP (red) epitope tag on Bod1 and the GST (green) epitope tag on Nuf2–Spc24. ( e ) Amount of bound protein in ( d ) was quantified relative to the input. Two asterisks indicate level of significance ( p

    Journal: Open Biology

    Article Title: The Ndc80 complex targets Bod1 to human mitotic kinetochores

    doi: 10.1098/rsob.170099

    Figure Lengend Snippet: The Ndc80 complex interacts with Bod1 in mitotic HeLa cell extracts and in vitro . ( a ) Graphic representation of the full-length Ndc80 complex. Domains of interest, including the microtubule and centromere binding regions of the complex, are labelled. ( b ) Co-precipitation of Ndc80 complex components Ndc80, Nuf2 and Spc24 from mitotic HeLa cell extracts with purified Bod1-GST. ( c ) Representation of the recombinant Ndc80 Bonsai complex (as described in [ 38 ]). Ndc80–Spc25 and GST–Nuf2–Spc24 are expressed as fusion proteins. Both fusion constructs are co-expressed in E. coli from a dual-expression vector. ( d ) Of note, 150 pmol recombinant Ndc80 Bonsai, consisting of dimers of one Ndc80–Spc25 fusion protein with one GST–Nuf2–Spc24 fusion protein, was immobilized on Sepharose beads and incubated with 1 nmol Bod1-MBP or MBP. Binding was allowed for 1 h. Proteins were resolved by SDS-PAGE and immunoblotted using simultaneous detection of the MBP (red) epitope tag on Bod1 and the GST (green) epitope tag on Nuf2–Spc24. ( e ) Amount of bound protein in ( d ) was quantified relative to the input. Two asterisks indicate level of significance ( p

    Article Snippet: Primary antibodies included mouse anti-B56α (1 : 500, Abcam), mouse anti-B56δ (1 : 500, Abcam), mouse anti-Ndc80 (1 : 1000, Abcam [9G3]), mouse anti-Nuf2 (1 : 1000, Abcam), rabbit anti-Spc24 (1 : 1000, Abcam [EPR11548(B)]), mouse anti-MBP (1 : 20 000, NEB), goat anti-GST (1 : 5000, Abcam), mouse anti-Vinculin (1 : 10 000, Abcam [SPM227]), mouse anti-GFP (1 : 1000, Roche), rabbit anti-Bod1 (1 : 500, Abcam), polyclonal sheep anti-Bod1 (2 µg ml−1 ).

    Techniques: In Vitro, Binding Assay, Purification, Recombinant, Construct, Expressing, Plasmid Preparation, Incubation, SDS Page

    Knockdown of Spc24 causes aneuploidy ( A ) Oocytes in MII stage were collected and chromosome spreads were performed in control and Spc24 RNAi oocytes. Scale bars: 20 μm. ( B ) The numbers of univalents in the oocytes in A a; b; c are 20; 19; 23, (“a” is control oocyte, “b,c” are RNAi oocytes), respectively. The total numbers of analyzed oocytes are indicated (n).

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Knockdown of Spc24 causes aneuploidy ( A ) Oocytes in MII stage were collected and chromosome spreads were performed in control and Spc24 RNAi oocytes. Scale bars: 20 μm. ( B ) The numbers of univalents in the oocytes in A a; b; c are 20; 19; 23, (“a” is control oocyte, “b,c” are RNAi oocytes), respectively. The total numbers of analyzed oocytes are indicated (n).

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques:

    Knockdown of Spc24 accelerates polar body extrusion ( A ) Levels of Spc24 mRNA or protein in siRNA injected oocytes. The GV stage oocytes were microinjected with negative control siRNA or Spc24 siRNA and incubated for 24 h in M2 medium containing 200 μM IBMX before collecting the oocytes for real time quantitative PCR or western blot. ( B ) Timing of polar body extrusion was determined in Spc24-depleted oocytes and control oocytes. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Knockdown of Spc24 accelerates polar body extrusion ( A ) Levels of Spc24 mRNA or protein in siRNA injected oocytes. The GV stage oocytes were microinjected with negative control siRNA or Spc24 siRNA and incubated for 24 h in M2 medium containing 200 μM IBMX before collecting the oocytes for real time quantitative PCR or western blot. ( B ) Timing of polar body extrusion was determined in Spc24-depleted oocytes and control oocytes. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques: Injection, Negative Control, Incubation, Real-time Polymerase Chain Reaction, Western Blot

    Mad2-mediated SAC inactivation in Spc24-depleted oocytes Control and Spc24-depleted oocytes were fixed at 5 h following release from IBMX. ( A ) Oocytes were immunostained with anti-Mad2 antibody (green) and Hoechst 33342 (red). Scale bars: 20 μm. ( B ) Oocytes were immunostained with anti-Bub3 antibody (green) and Hoechst 33342 (red). Scale bars: 20 μm. Quantification of fluorescent intensity of Mad2 or Bub3 is shown in the right panel of images, respectively. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Mad2-mediated SAC inactivation in Spc24-depleted oocytes Control and Spc24-depleted oocytes were fixed at 5 h following release from IBMX. ( A ) Oocytes were immunostained with anti-Mad2 antibody (green) and Hoechst 33342 (red). Scale bars: 20 μm. ( B ) Oocytes were immunostained with anti-Bub3 antibody (green) and Hoechst 33342 (red). Scale bars: 20 μm. Quantification of fluorescent intensity of Mad2 or Bub3 is shown in the right panel of images, respectively. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques:

    Loss of Spc24 causes misaligned chromosomes in meiotic oocytes ( A ) Abnormal chromosome alignment in MII oocytes after microinjection of Spc24 siRNA. In the control group, most oocytes showed normal chromosome alignment, while in the Spc24-depleted oocytes, most oocytes showed severely misaligned chromosomes. α-tubulin (green); DNA (red). Scale bars: 20 μm. ( B ) The rates of oocytes with misaligned chromosomes in the siRNA injection and control group. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Loss of Spc24 causes misaligned chromosomes in meiotic oocytes ( A ) Abnormal chromosome alignment in MII oocytes after microinjection of Spc24 siRNA. In the control group, most oocytes showed normal chromosome alignment, while in the Spc24-depleted oocytes, most oocytes showed severely misaligned chromosomes. α-tubulin (green); DNA (red). Scale bars: 20 μm. ( B ) The rates of oocytes with misaligned chromosomes in the siRNA injection and control group. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques: Injection

    Correct kinetochore-microtubule attachment depends on Spc24 ( A ) Control and Spc24-depleted MI oocytes were cold-treated and stained with anti-tubulin (microtubules, green), ACA (kinetochores, purple), and Hoechst 33342 (chromosomes, blue). Enlarged views for the kinetochore-microtubule attachments in the right panel. Aa: normal attachment; Ab-e: loss of attachment. Scale bars: 10 μm. ( B ) Quantitative analysis of kinetochore-microtubule attachments in control and Spc24-depleted oocytes. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Correct kinetochore-microtubule attachment depends on Spc24 ( A ) Control and Spc24-depleted MI oocytes were cold-treated and stained with anti-tubulin (microtubules, green), ACA (kinetochores, purple), and Hoechst 33342 (chromosomes, blue). Enlarged views for the kinetochore-microtubule attachments in the right panel. Aa: normal attachment; Ab-e: loss of attachment. Scale bars: 10 μm. ( B ) Quantitative analysis of kinetochore-microtubule attachments in control and Spc24-depleted oocytes. Data are expressed as mean ± SEM of at least 3 independent experiments. *Significantly different ( P

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques: Staining

    Subcellular localization of Spc24 during mouse oocyte meiotic maturation Oocytes were collected after culture for 0, 4, 8 and 12 h, the time points when most oocytes had reached the GV, GVBD, MI and MII stages, respectively. ( A ) Confocal microcopy showing the subcellular localization of Spc24 (red) in mouse oocytes at GV, GVBD, MI and MII stages. DNA (blue) was counterstained with Hoechst 33342. Scale bars: 20 μm. ( B ) The localization of Spc24 (green) and ACA (purple) on chromosome spreads at MI and MII stages. DNA (blue) was counterstained with Hoechst 33342. Scale bars: 20 μm.

    Journal: Oncotarget

    Article Title: Spc24 is required for meiotic kinetochore-microtubule attachment and production of euploid eggs

    doi: 10.18632/oncotarget.12453

    Figure Lengend Snippet: Subcellular localization of Spc24 during mouse oocyte meiotic maturation Oocytes were collected after culture for 0, 4, 8 and 12 h, the time points when most oocytes had reached the GV, GVBD, MI and MII stages, respectively. ( A ) Confocal microcopy showing the subcellular localization of Spc24 (red) in mouse oocytes at GV, GVBD, MI and MII stages. DNA (blue) was counterstained with Hoechst 33342. Scale bars: 20 μm. ( B ) The localization of Spc24 (green) and ACA (purple) on chromosome spreads at MI and MII stages. DNA (blue) was counterstained with Hoechst 33342. Scale bars: 20 μm.

    Article Snippet: For staining of Spc24 or Mad2 or Bub3, oocytes were incubated overnight at 4°C with anti-Spc24 antibody (1:100); anti-Mad2 antibody (1:20); anti-Bub3 antibody (1:50), respectively.

    Techniques:

    SPC24 knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: SPC24 knockdown inhibits osteosarcoma xenograft tumor growth in nude mice model (A-B) Representative western blot analysis showing SPC24 expression in shN and shSPC24 transfected 143b and U2OS cells subcutaneously injected into 5-week old nude mice. (B) The tumor weights of xenograft tumors derived from control and SPC24 knockdown 143B and U2OS cells. Note: Values represent mean ± SEM; ** denotes p

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Mouse Assay, Western Blot, Expressing, Transfection, Injection, Derivative Assay

    SPC24 knockdown promotes apoptosis and G1-S cell cycle arrest in 143B and U2OS cells (A) Flow cytometry analysis of AnnexinV-FITC and propidium iodide stained control (siN) and SPC24 knockdown 143B and U2OS cells. AnnexinV + PI − cells and AnnexinV + PI + cells represent early and late apoptotic cells, respectively. (B) Representative western blot showing SPC24 and PARP/cleaved PARP levels in control and siSPC24 transfected OS cells at 72 h. (C) Flow cytometry analysis of cell cycle distribution in control (siN) and SPC24 knockdown 143B and U2OS cells. Cells were stained with propidium iodide.

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: SPC24 knockdown promotes apoptosis and G1-S cell cycle arrest in 143B and U2OS cells (A) Flow cytometry analysis of AnnexinV-FITC and propidium iodide stained control (siN) and SPC24 knockdown 143B and U2OS cells. AnnexinV + PI − cells and AnnexinV + PI + cells represent early and late apoptotic cells, respectively. (B) Representative western blot showing SPC24 and PARP/cleaved PARP levels in control and siSPC24 transfected OS cells at 72 h. (C) Flow cytometry analysis of cell cycle distribution in control (siN) and SPC24 knockdown 143B and U2OS cells. Cells were stained with propidium iodide.

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Flow Cytometry, Cytometry, Staining, Western Blot, Transfection

    SPC24 knockdown inhibits EGFR/Ras/ERK signaling in 143B and U2OS cells (A-B) Representative western blot showing SPC24, EGFR, K-Ras and p-ERK levels in siN or siSPC24 transfected (A) 143B and (B) U2OS cells. (C-D) Representative western blot showing SPC24, EGFR, K-Ras and p-ERK levels in siN or siEGFR transfected (A) 143B and (B) U2OS cells.

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: SPC24 knockdown inhibits EGFR/Ras/ERK signaling in 143B and U2OS cells (A-B) Representative western blot showing SPC24, EGFR, K-Ras and p-ERK levels in siN or siSPC24 transfected (A) 143B and (B) U2OS cells. (C-D) Representative western blot showing SPC24, EGFR, K-Ras and p-ERK levels in siN or siEGFR transfected (A) 143B and (B) U2OS cells.

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Western Blot, Transfection

    SPC24 knockdown increases invasiveness and EMT in 143B and U2OS cells (A-B) Representative images (250X) showing transwell invasion assay results of siN or siSPC24 transfected 143B and U2OS cells. (C-D) Representative western blots showing E-cadherin and SPC24 levels in siN or siSPC24 transfected 143B and U2OS cells at 72 h.

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: SPC24 knockdown increases invasiveness and EMT in 143B and U2OS cells (A-B) Representative images (250X) showing transwell invasion assay results of siN or siSPC24 transfected 143B and U2OS cells. (C-D) Representative western blots showing E-cadherin and SPC24 levels in siN or siSPC24 transfected 143B and U2OS cells at 72 h.

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Transwell Invasion Assay, Transfection, Western Blot

    Human osteosarcoma patient tissues show increased SPC24 and p-ERK and decreased E-cadherin levels (A) Representative western blot shows SPC24 levels in normal and OS patient tissue samples. B-actin was used as control. (B) QRT-PCR analysis of SPC24 mRNA levels in normal and OS patient tissue samples. (C) Representative images (magnification: 20x) show immunohistochemical staining of SPC24, p-ERK and E-cadherin in normal and OS tissue samples (Scale bar, 125μm). (D) Semiquantitative analysis of SPC24, p-ERK and E-cadherin expression in normal and OS cancer tissue samples based on immunohistochemical staining.

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: Human osteosarcoma patient tissues show increased SPC24 and p-ERK and decreased E-cadherin levels (A) Representative western blot shows SPC24 levels in normal and OS patient tissue samples. B-actin was used as control. (B) QRT-PCR analysis of SPC24 mRNA levels in normal and OS patient tissue samples. (C) Representative images (magnification: 20x) show immunohistochemical staining of SPC24, p-ERK and E-cadherin in normal and OS tissue samples (Scale bar, 125μm). (D) Semiquantitative analysis of SPC24, p-ERK and E-cadherin expression in normal and OS cancer tissue samples based on immunohistochemical staining.

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Western Blot, Quantitative RT-PCR, Immunohistochemistry, Staining, Expressing

    SPC24 knockdown decreases growth, viability and colony formation in 143B and U2OS cells (A-B) Representative western blot images showing SPC24 expression in siSPC24 and siN transfected (A) 143B and (B) U2OS cells. β-actin is the loading control. (C-D) MTT assay showing viability of siSPC24 and siN transfected (C) 143B and (D) U2OS cells at 24, 48 and 72 h. (E-F) Soft-agar colony formation assay showing crystal violet stained colonies from siSPC24 and siN transfected (E) 143B and (F) U2OS cells. Note: * denotes p

    Journal: Oncotarget

    Article Title: SPC24 promotes osteosarcoma progression by increasing EGFR/MAPK signaling

    doi: 10.18632/oncotarget.22167

    Figure Lengend Snippet: SPC24 knockdown decreases growth, viability and colony formation in 143B and U2OS cells (A-B) Representative western blot images showing SPC24 expression in siSPC24 and siN transfected (A) 143B and (B) U2OS cells. β-actin is the loading control. (C-D) MTT assay showing viability of siSPC24 and siN transfected (C) 143B and (D) U2OS cells at 24, 48 and 72 h. (E-F) Soft-agar colony formation assay showing crystal violet stained colonies from siSPC24 and siN transfected (E) 143B and (F) U2OS cells. Note: * denotes p

    Article Snippet: SPC24 knockdown in U2OS and 143B cell lines The stable SPC24 knockout U20S and 143B cell lines were generated by transfecting retroviral shRNA vectors specific for SPC24 (OriGene, Rockville, MD).

    Techniques: Western Blot, Expressing, Transfection, MTT Assay, Soft Agar Assay, Staining