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  • 99
    Thermo Fisher sigenome smartpool sirna
    Sigenome Smartpool Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sigenome smartpool sirna
    Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control <t>siRNA</t> to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b <t>siGENOME</t> <t>SMARTpool</t> oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p
    Sigenome Smartpool Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 90/100, based on 246 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sigenome smartpool sirna reagents
    Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control <t>siRNA</t> to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b <t>siGENOME</t> <t>SMARTpool</t> oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p
    Sigenome Smartpool Sirna Reagents, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher sigenome smartpool sirnas
    Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control <t>siRNA</t> to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b <t>siGENOME</t> <t>SMARTpool</t> oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p
    Sigenome Smartpool Sirnas, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery human sigenome smartpool sirna libraries
    Primary Screen. HUVEC cells were transfected with 20 pmol of the combined Human siARRAY® <t>SMARTpool®</t> <t>siRNA</t> Libraries for Kinases (targeting 779 genes) and Cell Cycle Regulators (targeting 111 genes). Cell viability was analyzed 72 hrs post-Nucleofection®. (A) Representation of robust Z-scores of cell viability measures from 1 screening experiment. (B-E) Robust Z-scores of all primary hits with MAD of > 3 in all three (B), two of three (C), or one of three (D) independent experiments and with MAD of
    Human Sigenome Smartpool Sirna Libraries, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sirna2 smartpool sigenome fmnl2 sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Sirna2 Smartpool Sigenome Fmnl2 Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery gene specific sigenome smartpool sirnas
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Gene Specific Sigenome Smartpool Sirnas, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transfection mouse sam68 sigenome smartpool sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Transfection Mouse Sam68 Sigenome Smartpool Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery transfection sigenome smartpool sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Transfection Sigenome Smartpool Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery mouse egfr sigenome smartpool sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Mouse Egfr Sigenome Smartpool Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery mouse sigenome sirna library smartpool
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Mouse Sigenome Sirna Library Smartpool, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery double stranded sigenome smartpool oligonucleotides sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Double Stranded Sigenome Smartpool Oligonucleotides Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sigenome sirna reagents human smartpool sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Sigenome Sirna Reagents Human Smartpool Sirna, supplied by Horizon Discovery, 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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    Horizon Discovery sigenome sirna smartpool custom library
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Sigenome Sirna Smartpool Custom Library, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sigenome sirna smartpool reagents against pdk2
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
    Sigenome Sirna Smartpool Reagents Against Pdk2, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery sigenome on targetplus smartpool sirna
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
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    Horizon Discovery sigenome rtf smartpool sirna library
    <t>FMNL2</t> knockdown modifies human trabecular meshwork cell morphology. Following <t>siRNA-mediated</t> knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and <t>-siRNA2</t> caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P
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    Image Search Results


    Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control siRNA to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b siGENOME SMARTpool oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p

    Journal: Breast Cancer Research : BCR

    Article Title: A novel role for signal transducer and activator of transcription 5b (STAT5b) in ?1-integrin-mediated human breast cancer cell migration

    doi: 10.1186/bcr2341

    Figure Lengend Snippet: Expression of wild-type, Y699F-, or dominant-negative STAT5b rescues migration, but expression of R618K-STAT5b does not. (a) Domain structure of the STAT5b protein depicting the amino terminus (N-term), DNA-binding domain (DBD), Src homology 2 domain (SH2), transactivation domain (TAD), and the location of the conserved tyrosine residue, Y699, and arginine residue, R618. (b) MDA-MB-231 breast cancer cells were transfected with control siRNA to luciferase (siLuc) or siRNA specific to STAT5b (siSTAT5b siGENOME SMARTpool oligonucleotide #3) alone or in the presence of HA-tagged wild-type (wt-STAT5b), Y699F (YF-STAT5b), dominant-negative (dn-STAT5b), or R618K (RK-STAT5b) STAT5b constructs engineered to be immune to siRNA knockdown. Seventy-two hours after transfection, trans-well assays were performed for 6 hours, as described in Figure 1. Results are graphed as relative migration, compared with siLuc control. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( p

    Article Snippet: siRNA Transfection BT-549 and MDA-MB-231 cells were transfected with siGENOME SMARTpool siRNA targeting human STAT5b or individual custom oligonucleotides specific for STAT5a or STAT5b (siGENOME STAT5b SMARTpool duplex #3), or luciferase duplex control, all purchased from Dharmacon (Lafayette, CO).

    Techniques: Expressing, Dominant Negative Mutation, Migration, Binding Assay, Multiple Displacement Amplification, Transfection, Luciferase, Construct

    STAT5b knockdown inhibits breast cancer cell migration. (a) BT-549 or MDA-MB-231 breast cancer cells were transfected with no siRNA (con), control siRNA for luciferase (siLuc), siSTAT5b SMARTpool (BT-549 cells), or siSTAT5b SMARTpool duplex #3 (MDA-MB-231 cells). Seventy-two hours after transfection, cells were plated in serum-free media in trans-well chambers. Media containing 1% fetal bovine serum (FBS; BT-549) or 10% FBS (MDA-MB-231) were placed in the lower chambers. After 3 hours (BT-549) or 6 hours (MDA-MB-231), cells were fixed, stained with crystal violet, and the number of migratory cells was counted. Results are graphed as the average number of migratory cells per field ± SEM. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( P

    Journal: Breast Cancer Research : BCR

    Article Title: A novel role for signal transducer and activator of transcription 5b (STAT5b) in ?1-integrin-mediated human breast cancer cell migration

    doi: 10.1186/bcr2341

    Figure Lengend Snippet: STAT5b knockdown inhibits breast cancer cell migration. (a) BT-549 or MDA-MB-231 breast cancer cells were transfected with no siRNA (con), control siRNA for luciferase (siLuc), siSTAT5b SMARTpool (BT-549 cells), or siSTAT5b SMARTpool duplex #3 (MDA-MB-231 cells). Seventy-two hours after transfection, cells were plated in serum-free media in trans-well chambers. Media containing 1% fetal bovine serum (FBS; BT-549) or 10% FBS (MDA-MB-231) were placed in the lower chambers. After 3 hours (BT-549) or 6 hours (MDA-MB-231), cells were fixed, stained with crystal violet, and the number of migratory cells was counted. Results are graphed as the average number of migratory cells per field ± SEM. One-way ANOVA with Tukey's post-test was used to determine statistical significance ( P

    Article Snippet: siRNA Transfection BT-549 and MDA-MB-231 cells were transfected with siGENOME SMARTpool siRNA targeting human STAT5b or individual custom oligonucleotides specific for STAT5a or STAT5b (siGENOME STAT5b SMARTpool duplex #3), or luciferase duplex control, all purchased from Dharmacon (Lafayette, CO).

    Techniques: Migration, Multiple Displacement Amplification, Transfection, Luciferase, Staining

    CD44 loss and apoptosis in bladder cancer cells +/− AGL. a UMUC3 shCTL and shAGL cells were plated and 24 h later transfected with scrambled siRNA (siCTL) or siGENOME SMARTpool siRNA against CD44(siCD44). Details of siRNA are in Material and Methods . Cells were lysed 48 h after transfection and Western blot was carried out for proteins involved in apoptosis. b Densitometric analysis of cleaved apoptotic proteins normalized to total protein and the UMUC3 shCTL siCTL sample ( n = 3). DR5 and Fas normalized to Actin and the UMUC3 shCTL siCTL sample ( n = 3). Results are shown as mean ± SD, * P

    Journal: BMC Cancer

    Article Title: CD44 and RHAMM are essential for rapid growth of bladder cancer driven by loss of Glycogen Debranching Enzyme (AGL)

    doi: 10.1186/s12885-016-2756-5

    Figure Lengend Snippet: CD44 loss and apoptosis in bladder cancer cells +/− AGL. a UMUC3 shCTL and shAGL cells were plated and 24 h later transfected with scrambled siRNA (siCTL) or siGENOME SMARTpool siRNA against CD44(siCD44). Details of siRNA are in Material and Methods . Cells were lysed 48 h after transfection and Western blot was carried out for proteins involved in apoptosis. b Densitometric analysis of cleaved apoptotic proteins normalized to total protein and the UMUC3 shCTL siCTL sample ( n = 3). DR5 and Fas normalized to Actin and the UMUC3 shCTL siCTL sample ( n = 3). Results are shown as mean ± SD, * P

    Article Snippet: HA (cat. # GLR001) was obtained from R & D systems (Minneapolis, MN). siRNA sequences 5’-GGTTTGTGATTCAGACACT-3’ was used at a concentration of 50nM to knockdown HAS2 (siHAS2) as previously reported [ ]. siGENOME SMARTpool siRNAs were used to knockdown CD44 (M-009999-03-0005, siCD44) and RHAMM (M-010409-01-0005, siRHAMM) at a concentration of 20nM. siRNA’s were purchased from Dharmacon (Lafayette, CO) and transfected using Lipofectamine RNAiMAX (Invitrogen) using manufacturer instructions.

    Techniques: Transfection, Western Blot

    RHAMM loss and apoptosis in bladder cancer cells +/− AGL. a UMUC3 shCTL and shAGL cells were plated and 24 h later transfected with scrambled siRNA (siCTL) or siGENOME SMARTpool siRNA against RHAMM (siRHAMM). Details of siRNA are in Material and Methods . Cells were lysed 48 h after transfection and Western blot was carried out for proteins involved in apoptosis. b Densitometric analysis of cleaved apoptotic proteins normalized to total protein and the UMUC3 shCTL siCTL sample ( n = 3). DR5 and Fas normalized to Actin and the UMUC3 shCTL siCTL sample ( n = 3). Results are shown as mean ± SD, * P

    Journal: BMC Cancer

    Article Title: CD44 and RHAMM are essential for rapid growth of bladder cancer driven by loss of Glycogen Debranching Enzyme (AGL)

    doi: 10.1186/s12885-016-2756-5

    Figure Lengend Snippet: RHAMM loss and apoptosis in bladder cancer cells +/− AGL. a UMUC3 shCTL and shAGL cells were plated and 24 h later transfected with scrambled siRNA (siCTL) or siGENOME SMARTpool siRNA against RHAMM (siRHAMM). Details of siRNA are in Material and Methods . Cells were lysed 48 h after transfection and Western blot was carried out for proteins involved in apoptosis. b Densitometric analysis of cleaved apoptotic proteins normalized to total protein and the UMUC3 shCTL siCTL sample ( n = 3). DR5 and Fas normalized to Actin and the UMUC3 shCTL siCTL sample ( n = 3). Results are shown as mean ± SD, * P

    Article Snippet: HA (cat. # GLR001) was obtained from R & D systems (Minneapolis, MN). siRNA sequences 5’-GGTTTGTGATTCAGACACT-3’ was used at a concentration of 50nM to knockdown HAS2 (siHAS2) as previously reported [ ]. siGENOME SMARTpool siRNAs were used to knockdown CD44 (M-009999-03-0005, siCD44) and RHAMM (M-010409-01-0005, siRHAMM) at a concentration of 20nM. siRNA’s were purchased from Dharmacon (Lafayette, CO) and transfected using Lipofectamine RNAiMAX (Invitrogen) using manufacturer instructions.

    Techniques: Transfection, Western Blot

    BUB1B knockdown inhibits anchorage-independent growth of lung adenocarcinoma cell lines A. Soft agar colony formation of mouse lung adenocarcinoma cell lines transfected with siNTC or mouse Bub1b siRNA SMARTpool. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from two independent experiments. The t-test p-values for siNTC vs siBub1b in LKR13 and LKR13_shTP53 cells were 0.09 and 0.07, respectively. B. BUB1B protein levels in LKPH2 cells expressing inducible shNTC or shBub1b 72 hours after induction with 100ng/ml doxycycline. C. Soft agar colony formation of LKPH2 cells from B. Colony numbers were shown as relative values normalized to no doxycycline treatment controls. Mean±SEM was calculated from three independent experiments. D. Cell proliferation of LKPH2 cells from B measured by CellTiterGlo. Values were normalized to no doxycycline treatment controls. Mean±SEM was calculated from two independent experiments. E. Soft agar colony formation of human lung adenocarcinoma cell lines transfected with siNTC or human BUB1B siRNA SMARTpool. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from two (HCC827, H23 and H460) to three independent experiments. The t-test p-values for siNTC vs siBub1b in HCC827, H23 and H460 cells were 0.1, 0.09 and 0.07, respectively. WT/wt, wild type. MT/mt, mutant. F and G. Cell lines in E were grouped based on their KRAS (F) or TP53 (G) status. *** indicates p

    Journal: Genes & Cancer

    Article Title: Requirement for BUB1B/BUBR1 in tumor progression of lung adenocarcinoma

    doi:

    Figure Lengend Snippet: BUB1B knockdown inhibits anchorage-independent growth of lung adenocarcinoma cell lines A. Soft agar colony formation of mouse lung adenocarcinoma cell lines transfected with siNTC or mouse Bub1b siRNA SMARTpool. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from two independent experiments. The t-test p-values for siNTC vs siBub1b in LKR13 and LKR13_shTP53 cells were 0.09 and 0.07, respectively. B. BUB1B protein levels in LKPH2 cells expressing inducible shNTC or shBub1b 72 hours after induction with 100ng/ml doxycycline. C. Soft agar colony formation of LKPH2 cells from B. Colony numbers were shown as relative values normalized to no doxycycline treatment controls. Mean±SEM was calculated from three independent experiments. D. Cell proliferation of LKPH2 cells from B measured by CellTiterGlo. Values were normalized to no doxycycline treatment controls. Mean±SEM was calculated from two independent experiments. E. Soft agar colony formation of human lung adenocarcinoma cell lines transfected with siNTC or human BUB1B siRNA SMARTpool. Colony numbers were shown as relative values normalized to siNTC controls. Mean±SEM was calculated from two (HCC827, H23 and H460) to three independent experiments. The t-test p-values for siNTC vs siBub1b in HCC827, H23 and H460 cells were 0.1, 0.09 and 0.07, respectively. WT/wt, wild type. MT/mt, mutant. F and G. Cell lines in E were grouped based on their KRAS (F) or TP53 (G) status. *** indicates p

    Article Snippet: Mouse kinome siRNA screen and hit prioritization LKR13_shTP53 cells were transfected in quintuplicate in 96-well plates with the Dharmacon siGENOME SMARTpool siRNA library targeting 713 mouse kinases.

    Techniques: Transfection, Expressing, Mutagenesis

    Primary Screen. HUVEC cells were transfected with 20 pmol of the combined Human siARRAY® SMARTpool® siRNA Libraries for Kinases (targeting 779 genes) and Cell Cycle Regulators (targeting 111 genes). Cell viability was analyzed 72 hrs post-Nucleofection®. (A) Representation of robust Z-scores of cell viability measures from 1 screening experiment. (B-E) Robust Z-scores of all primary hits with MAD of > 3 in all three (B), two of three (C), or one of three (D) independent experiments and with MAD of

    Journal: Journal of RNAi and Gene Silencing : An International Journal of RNA and Gene Targeting Research

    Article Title: First siRNA library screening in hard-to-transfect HUVEC cells

    doi:

    Figure Lengend Snippet: Primary Screen. HUVEC cells were transfected with 20 pmol of the combined Human siARRAY® SMARTpool® siRNA Libraries for Kinases (targeting 779 genes) and Cell Cycle Regulators (targeting 111 genes). Cell viability was analyzed 72 hrs post-Nucleofection®. (A) Representation of robust Z-scores of cell viability measures from 1 screening experiment. (B-E) Robust Z-scores of all primary hits with MAD of > 3 in all three (B), two of three (C), or one of three (D) independent experiments and with MAD of

    Article Snippet: MATERIALS AND METHODS The siRNA reagents used were Dharmacon Human siGENOME® SMARTpool® siRNA Libraries for Protein Kinases (targeting 779 genes) and Cell Cycle Regulation (targeting 111 genes) (Thermo Fisher Scientific).

    Techniques: Transfection

    Determination of optimal assay conditions. In three independent experiments, HUVEC cells were transfected with 20 pmol SMARTpool® siRNA targeting PLK-1 (A, B) or CHEK-1 (B, C) and siGENOME® non-targeting control. Cell viability was analyzed at different time points post Nucleofection® (A/B: 24, 48, 72 and 96 hrs; C: 72 hrs). Values were normalized to the negative control samples (A, B) or to untreated cells (C). The rightmost dots in C represents the mean and SD of the 60 individual values.

    Journal: Journal of RNAi and Gene Silencing : An International Journal of RNA and Gene Targeting Research

    Article Title: First siRNA library screening in hard-to-transfect HUVEC cells

    doi:

    Figure Lengend Snippet: Determination of optimal assay conditions. In three independent experiments, HUVEC cells were transfected with 20 pmol SMARTpool® siRNA targeting PLK-1 (A, B) or CHEK-1 (B, C) and siGENOME® non-targeting control. Cell viability was analyzed at different time points post Nucleofection® (A/B: 24, 48, 72 and 96 hrs; C: 72 hrs). Values were normalized to the negative control samples (A, B) or to untreated cells (C). The rightmost dots in C represents the mean and SD of the 60 individual values.

    Article Snippet: MATERIALS AND METHODS The siRNA reagents used were Dharmacon Human siGENOME® SMARTpool® siRNA Libraries for Protein Kinases (targeting 779 genes) and Cell Cycle Regulation (targeting 111 genes) (Thermo Fisher Scientific).

    Techniques: Transfection, Negative Control

    Hit validation. HUVEC cells were transfected with 20 pmol (if not indicated differently) siGENOME® (siG) SMARTpool® or single siRNA #1 - 4 (from the de-convoluted pool) targeting CDK4, COPB2, MYC or PYCS. CHEK-1 and siGENOME® Non-Targeting siRNA #1 (control siRNA) and siRNA targeting CHEK-1 (CHEK-1) served as controls. 72 hrs post- Nucleofection® cell viability was analyzed and normalized to control the siRNA (A, C) and mRNA levels were determined for CDK4 (B) and COPB2 (B, C) and normalized to cyclophilin B mRNA and the control siRNA.

    Journal: Journal of RNAi and Gene Silencing : An International Journal of RNA and Gene Targeting Research

    Article Title: First siRNA library screening in hard-to-transfect HUVEC cells

    doi:

    Figure Lengend Snippet: Hit validation. HUVEC cells were transfected with 20 pmol (if not indicated differently) siGENOME® (siG) SMARTpool® or single siRNA #1 - 4 (from the de-convoluted pool) targeting CDK4, COPB2, MYC or PYCS. CHEK-1 and siGENOME® Non-Targeting siRNA #1 (control siRNA) and siRNA targeting CHEK-1 (CHEK-1) served as controls. 72 hrs post- Nucleofection® cell viability was analyzed and normalized to control the siRNA (A, C) and mRNA levels were determined for CDK4 (B) and COPB2 (B, C) and normalized to cyclophilin B mRNA and the control siRNA.

    Article Snippet: MATERIALS AND METHODS The siRNA reagents used were Dharmacon Human siGENOME® SMARTpool® siRNA Libraries for Protein Kinases (targeting 779 genes) and Cell Cycle Regulation (targeting 111 genes) (Thermo Fisher Scientific).

    Techniques: Transfection

    FMNL2 knockdown modifies human trabecular meshwork cell morphology. Following siRNA-mediated knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and -siRNA2 caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P

    Journal: Nature Communications

    Article Title: A multiethnic genome-wide association study of primary open-angle glaucoma identifies novel risk loci

    doi: 10.1038/s41467-018-04555-4

    Figure Lengend Snippet: FMNL2 knockdown modifies human trabecular meshwork cell morphology. Following siRNA-mediated knockdown of FMNL2 for 24 h, HTM cells were serum-starved for 24 h in order to disrupt the normal organization of actin fibers. The cells were then supplemented with fetal bovine serum (FBS) for 10 min or 1 h to induce formation of actin stress fibers and cell spreading. a Both the FMNL2 siRNA1 and -siRNA2 caused significant knockdown of FMNL2 mRNA as assessed by qPCR. The expression of FMNL3 , a paralog of FMNL2 was not altered. b Fluorescent microscopic images of phalloidin-FITC-stained F-actin shows that the actin stress fibers are poorly detectable in serum-starved control siRNA-transfected cells (−FBS), and following treatment with FBS (+FBS for 10 min or 1 h) the stress fibers are prominently visible. Consistent with appearance of actin stress fibers, the control siRNA-transfected HTM cells treated with FBS for 10 min are well spread. The actin stress fibers of the FMNL2 -siRNA1 transfected HTM cells are poorly formed even after 1 h treatment with FBS. Moreover, many of these cells appear deformed/rounded or in general exhibiting poor cell spreading. c A bar graph showing proportion of cells (show as percentage) in each of the three groups of cells with distinct morphology (see Methods). In presence of FBS, the percentage of deformed/rounded or modestly spread cells (intermediate) is significantly more in FMNL2 siRNA1 transfected as compared to control siRNA-transfected HTM cells, which primarily show a well-spread morphology. d Magnified fluorescent miscroscopic images of phalloidin-FITC-stained F-actin presented in b . White arrows indicate cells with deformed/rounded morphology, arrowheads indicate modestly spread cells (intermediate), and asterisks indicate well-spread cells. Results are mean ± s.e.m. of three independent experiments. Significance of difference was determined by unpaired two-tailed t -test and Pearson’ Chi-squared test for a and c , respectively. *** P

    Article Snippet: For gene silencing, we used two separate siRNAs targeting different regions of FMNL2 . siRNA1: SMARTpool ON-TARGETplus FMNL2 siRNA (Dharmacon, #Cat L-031993-01). siRNA2: SMARTpool siGENOME FMNL2 siRNA (Dharmacon #Cat M-031993-01).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Staining, Transfection, Two Tailed Test