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OriGene flag sam68
Flag Sam68, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+recombinant+khdrbs1/SAM68+(KHDRBS1)+(NM_006559)+Human+Recombinant+Protein/pm39478550-111-10-12
Average 92 stars, based on 3 article reviews

flag sam68 - by Bioz Stars, 2026-10

92/100 stars

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Article Title: Intronic tRNAs of mitochondrial origin regulate constitutive and alternative splicing.
Article Snippet: .. Human recombinant KHDRBS1 (Sam68; CAT#: TP300263) was ordered from Origene (Rockville, Maryland, USA). ..

Article Title: Intronic tRNAs of mitochondrial origin regulate constitutive and alternative splicing
Article Snippet: .. Human recombinant KHDRBS1 (Sam68; CAT#: TP300263) was ordered from Origene (Rockville, Maryland, USA). ..

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Fig. 6 NimtRNA protein interaction analysis. a Biotinylated transcripts containing or lacking nimtRNAs were incubated with nuclear protein extracts. Subsequently, binding proteins were isolated and analyzed by PAGE; differential band patterns were excised and analyzed by MS. b GO analysis of nimtRNA transcript interacting proteins. c The top 10 nimtRNA transcript interacting proteins ranked by absolute abundance. d–f An electrophoretic mobility shift assay was performed with increasing concentrations of KHDRBS1 being incubated with a transcript of the nimtRNATyr (d), nimtRNATyr delT (e), or snoRNA SNORD115 (f). The unbound transcript is indicated by an asterisk; the transcript-protein complexes are indicated by triangles

Journal: Genome biology

Article Title: Intronic tRNAs of mitochondrial origin regulate constitutive and alternative splicing.

doi: 10.1186/s13059-020-02199-6

Figure Lengend Snippet: Fig. 6 NimtRNA protein interaction analysis. a Biotinylated transcripts containing or lacking nimtRNAs were incubated with nuclear protein extracts. Subsequently, binding proteins were isolated and analyzed by PAGE; differential band patterns were excised and analyzed by MS. b GO analysis of nimtRNA transcript interacting proteins. c The top 10 nimtRNA transcript interacting proteins ranked by absolute abundance. d–f An electrophoretic mobility shift assay was performed with increasing concentrations of KHDRBS1 being incubated with a transcript of the nimtRNATyr (d), nimtRNATyr delT (e), or snoRNA SNORD115 (f). The unbound transcript is indicated by an asterisk; the transcript-protein complexes are indicated by triangles

Article Snippet: Human recombinant KHDRBS1 (Sam68; CAT#: TP300263) was ordered from Origene (Rockville, Maryland, USA).

Techniques: Incubation, Binding Assay, Isolation, Electrophoretic Mobility Shift Assay

Figure 3. Sam68 Is an Essential Co-factor Mediating CWP Response in Human AML (A) mRNA expression proﬁle plot of CBP (CREBBP) in AML (red dots, n = 202) versus healthy (blue dots, n = 69) samples. Expression proﬁle is from Mills et al. (2009). (B) STRING analysis interaction network of gene products found to be (1) CBP direct interactors and (2) differentially expressed between AML and healthy donor samples with a p value %1.0 3 1010. CBP interactors with the strongest level of conﬁdence are illustrated. See also Key Resources Table. (C) mRNA expression proﬁle plot of Sam68 (KHDRBS1) in AML (red dots, n = 202) versus healthy (blue dots, n = 69) samples (p = 2.45 3 1014). (D) Western blot analyses of Sam68 protein expression in normal hematopoietic MNCs (from adult and cord blood) (n = 9) and AML patient blasts (n = 7). Samples used: #6, #7, and #8 (Table S2). Dotted lines indicate membrane cropping from original images presented in Figure S3A: Tracks #1 (MPB), #3 (CB MNCs), and #5 (CB Lin) were selected from Sam68 5-s exposure. Densitometry analysis of Sam68 signal versus GAPDH is expressed in the bar graph (***p < 0.0001). (E) Effect of CWP and ICG-001 on siRNA knockdowns of CBP and Sam68 in AML cells. Cell viability assessments were performed in knockdowns and controls 48 hr post transfection and 24 hr post drug treatment. Viability values are presented as relative to vehicle-treated control siRNA (vehicle + siCTRL, n = 11; CWP + siCTRL, n = 11; ICG-001 + siCTRL, n = 3; vehicle + siCBP, n = 6; CWP + siCBP, n = 6; ICG-001 + siCBP, n = 3; vehicle + siSam68, n = 8; CWP + siSam68, n = 8; ICG-001 + siSam68, n = 3; *p = 0.0346, ***p < 0.001). Error bars represent the SEM.

Journal: Cell chemical biology

Article Title: Sam68 Allows Selective Targeting of Human Cancer Stem Cells.

doi: 10.1016/j.chembiol.2017.05.026

Figure Lengend Snippet: Figure 3. Sam68 Is an Essential Co-factor Mediating CWP Response in Human AML (A) mRNA expression proﬁle plot of CBP (CREBBP) in AML (red dots, n = 202) versus healthy (blue dots, n = 69) samples. Expression proﬁle is from Mills et al. (2009). (B) STRING analysis interaction network of gene products found to be (1) CBP direct interactors and (2) differentially expressed between AML and healthy donor samples with a p value %1.0 3 1010. CBP interactors with the strongest level of conﬁdence are illustrated. See also Key Resources Table. (C) mRNA expression proﬁle plot of Sam68 (KHDRBS1) in AML (red dots, n = 202) versus healthy (blue dots, n = 69) samples (p = 2.45 3 1014). (D) Western blot analyses of Sam68 protein expression in normal hematopoietic MNCs (from adult and cord blood) (n = 9) and AML patient blasts (n = 7). Samples used: #6, #7, and #8 (Table S2). Dotted lines indicate membrane cropping from original images presented in Figure S3A: Tracks #1 (MPB), #3 (CB MNCs), and #5 (CB Lin) were selected from Sam68 5-s exposure. Densitometry analysis of Sam68 signal versus GAPDH is expressed in the bar graph (***p < 0.0001). (E) Effect of CWP and ICG-001 on siRNA knockdowns of CBP and Sam68 in AML cells. Cell viability assessments were performed in knockdowns and controls 48 hr post transfection and 24 hr post drug treatment. Viability values are presented as relative to vehicle-treated control siRNA (vehicle + siCTRL, n = 11; CWP + siCTRL, n = 11; ICG-001 + siCTRL, n = 3; vehicle + siCBP, n = 6; CWP + siCBP, n = 6; ICG-001 + siCBP, n = 3; vehicle + siSam68, n = 8; CWP + siSam68, n = 8; ICG-001 + siSam68, n = 3; *p = 0.0346, ***p < 0.001). Error bars represent the SEM.

Article Snippet: SUMOylated recombinant Sam68 (Origene TP300263) used as positive control was in vitro modified using a SUMOylation Kit (ENZO Life Sciences).

Techniques: Expressing, Western Blot, Membrane, Transfection, Control

Figure 4. Sam68 Is Associated with Selective Response to CWP in Human Breast and Colon CSC-like Models (A) CBP and Sam68 protein expression levels in normal versus neoplastic breast (normal adult dermal ﬁbroblasts [HDF-A], MCF-7, and MDA-MB-231: n = 3), and colon (normal intestinal progenitors [HIEC cells], SW480, and HT29: n = 3). Densitometry analyses were performed from the series of blots presented in Fig- ure S3B and expressed as relative amounts versus GAPDH loading control (*p % 0.019). See also Figures S4A and S4B. (B) Cell count-based selective toxicity assay on non-CSC MCF-7 and SW480 cells versus CSC-like MDA-MB-231 and HT29 lines comparing CWP (500 nM, 48 hr; breast n = 6, **p = 0.0062; colon n = 3, **p = 0.0019) and ICG-001 (3 mM, 48 hr; breast n = 4, **p = 0.0022; colon n = 3, *p = 0.0317). See also Figure S4C. (C) Ki67 expression proﬁling from vehicle and CWP-treated (500 nM, 48 hr) CSC-like cell lines (DMSO n = 4, CWP n = 8; ***p < 0.001). (D) Lentiviral-mediated overexpression of 6x-histidine-tagged full-length Sam68 in MCF-7 cells. Ratios of Hoechst-positive nuclei counts from vehicle versus CWP-treated (500 nM, 48 hr) cells are presented (n = 3, *p = 0.0345). EGFP-transduced MCF-7 cells were used as control. Error bars represent the SEM. See also Figures S4D and S4E.

Journal: Cell chemical biology

Article Title: Sam68 Allows Selective Targeting of Human Cancer Stem Cells.

doi: 10.1016/j.chembiol.2017.05.026

Figure Lengend Snippet: Figure 4. Sam68 Is Associated with Selective Response to CWP in Human Breast and Colon CSC-like Models (A) CBP and Sam68 protein expression levels in normal versus neoplastic breast (normal adult dermal ﬁbroblasts [HDF-A], MCF-7, and MDA-MB-231: n = 3), and colon (normal intestinal progenitors [HIEC cells], SW480, and HT29: n = 3). Densitometry analyses were performed from the series of blots presented in Fig- ure S3B and expressed as relative amounts versus GAPDH loading control (*p % 0.019). See also Figures S4A and S4B. (B) Cell count-based selective toxicity assay on non-CSC MCF-7 and SW480 cells versus CSC-like MDA-MB-231 and HT29 lines comparing CWP (500 nM, 48 hr; breast n = 6, **p = 0.0062; colon n = 3, **p = 0.0019) and ICG-001 (3 mM, 48 hr; breast n = 4, **p = 0.0022; colon n = 3, *p = 0.0317). See also Figure S4C. (C) Ki67 expression proﬁling from vehicle and CWP-treated (500 nM, 48 hr) CSC-like cell lines (DMSO n = 4, CWP n = 8; ***p < 0.001). (D) Lentiviral-mediated overexpression of 6x-histidine-tagged full-length Sam68 in MCF-7 cells. Ratios of Hoechst-positive nuclei counts from vehicle versus CWP-treated (500 nM, 48 hr) cells are presented (n = 3, *p = 0.0345). EGFP-transduced MCF-7 cells were used as control. Error bars represent the SEM. See also Figures S4D and S4E.

Article Snippet: SUMOylated recombinant Sam68 (Origene TP300263) used as positive control was in vitro modified using a SUMOylation Kit (ENZO Life Sciences).

Techniques: Expressing, Control, Cell Counting, Over Expression

Figure 5. CWP Treatment Selectively Affects Sam68 Cellular Distribution and CBP/b-Catenin Partnership in CSCs (A) CBP and Sam68 protein expression levels in normal versus neoplastic human pluripotent stem cells (PSCs and t-PSCs). GAPDH was used as loading control (p % 0.019). (B) Cell count-based selective toxicity assay for CWP and ICG-001 on human normal (n = 8) versus transformed (CWP, n = 3; ICG-001, n = 4) pluripotent stem cells (***p < 0.0001). (C) Sam68 immunostaining in vehicle control and CWP-treated PSCs (n = 8) and t-PSCs (n = 9). Scale bar, 50 mm. See also Figures S5H and S5J. (D) Quantiﬁcation of Sam68 nuclear protein levels upon CWP (100 nM, 48 hr) or ICG-001 (3 mM, 48 hr) treatment was performed by high-content imaging (***p % 0.0003). (E) Interaction assessment between Sam68, b-catenin, and CBP using co-immunoprecipitation (IP) upon CWP treatment in t-PSC (Sam68, n = 7; b-catenin, n = 5) and PSC (Sam68, n = 6; b-catenin, n = 3). See also Figure S5K. (F) Changes in CBP/Sam68 and CBP/b-catenin interaction levels were quantiﬁed in t-PSC and PSC models (**p % 0.01; n.s, not signiﬁcant). (G) Representative western blots of total H3K14ac (n = 3), H3K18ac (n = 6), and H3K4me3 (n = 3) levels in control and CWP-treated t-PSCs. Histone H3 and GAPDH were used as loading controls. (H) Quantiﬁcation of protein and histone modiﬁcation levels from western blots of control versus CWP-treated cells are represented in a bar chart (*p = 0.049; **p = 0.0098; ***p = 0.00012). Histone H3 and GAPDH were used as loading controls. (I) Co-immunoprecipitation (IP) kinetics assessing changes in interaction levels between CBP, b-catenin, and Sam68 in response to CWP (100 nM) in t-PSC. Error bars represent the SEM.

Journal: Cell chemical biology

Article Title: Sam68 Allows Selective Targeting of Human Cancer Stem Cells.

doi: 10.1016/j.chembiol.2017.05.026

Figure Lengend Snippet: Figure 5. CWP Treatment Selectively Affects Sam68 Cellular Distribution and CBP/b-Catenin Partnership in CSCs (A) CBP and Sam68 protein expression levels in normal versus neoplastic human pluripotent stem cells (PSCs and t-PSCs). GAPDH was used as loading control (p % 0.019). (B) Cell count-based selective toxicity assay for CWP and ICG-001 on human normal (n = 8) versus transformed (CWP, n = 3; ICG-001, n = 4) pluripotent stem cells (***p < 0.0001). (C) Sam68 immunostaining in vehicle control and CWP-treated PSCs (n = 8) and t-PSCs (n = 9). Scale bar, 50 mm. See also Figures S5H and S5J. (D) Quantiﬁcation of Sam68 nuclear protein levels upon CWP (100 nM, 48 hr) or ICG-001 (3 mM, 48 hr) treatment was performed by high-content imaging (***p % 0.0003). (E) Interaction assessment between Sam68, b-catenin, and CBP using co-immunoprecipitation (IP) upon CWP treatment in t-PSC (Sam68, n = 7; b-catenin, n = 5) and PSC (Sam68, n = 6; b-catenin, n = 3). See also Figure S5K. (F) Changes in CBP/Sam68 and CBP/b-catenin interaction levels were quantiﬁed in t-PSC and PSC models (**p % 0.01; n.s, not signiﬁcant). (G) Representative western blots of total H3K14ac (n = 3), H3K18ac (n = 6), and H3K4me3 (n = 3) levels in control and CWP-treated t-PSCs. Histone H3 and GAPDH were used as loading controls. (H) Quantiﬁcation of protein and histone modiﬁcation levels from western blots of control versus CWP-treated cells are represented in a bar chart (*p = 0.049; **p = 0.0098; ***p = 0.00012). Histone H3 and GAPDH were used as loading controls. (I) Co-immunoprecipitation (IP) kinetics assessing changes in interaction levels between CBP, b-catenin, and Sam68 in response to CWP (100 nM) in t-PSC. Error bars represent the SEM.

Article Snippet: SUMOylated recombinant Sam68 (Origene TP300263) used as positive control was in vitro modified using a SUMOylation Kit (ENZO Life Sciences).

Techniques: Expressing, Control, Cell Counting, Transformation Assay, Immunostaining, Imaging, Immunoprecipitation, Western Blot

Figure 6. SUMOylation of Sam68 Participates in CWP Response to CSCs (A) Representative immunoﬂuorescence images of Sam68 (red) and SUMO1 (green) distribution in control and CWP-treated (100 nM, 48 hr) t-PSC (n = 8). Hoechst (blue) was used as nuclear counterstaining. Magniﬁcation: 320. (B) Dot-blot estimation of global SUMOylation levels (total SUMO1) in human PSCs versus t-PSCs, and cord blood MNCs versus primary AML samples (n = 9). Samples used: #9, #10, and #11 (Table S2). (C) Immunodetection of SUMOylated Sam68 species in vehicle, CWP, and ICG-001 treated t-PSCs (88 kDa). Puriﬁed 6xHis-Sam68 in vitro SUMOylated was used as a positive control. GAPDH performed on input lysates was used as loading control. Quantitative analysis of t-PSCs (Sam68 immunoprecipitation [IP] + western blot [WB]: n = 3) and AML cells (total Sam68 WB: n = 3) showed increased levels of SUMOylated Sam68 in response to CWP (*p = 0.0214, #p = 0.0611). See also Figures S6A–S6C. (D) Schematic illustration of wild-type (WT) and semi-SUMOylable mutant (K96R) Sam68 constructs used for overexpression experiments in t-PSCs. (E) HCI assessment of Sam68 nuclear levels from GFP-positive populations for control vector (EGFP), as well as Sam68 WT and K96R mutant transduced t-PSCs treated with CWP (100 nM) or ICG-001 (3 mM). Data are expressed as relative amounts versus control EGFP for each treatment (n = 6, **p % 0.0094). Repre- sentative micrographs for CWP versus vehicle controls are also presented. Magniﬁcation: 320. (F) Cell counts of GFP-positive populations from t-PSCs transduced with control (EGFP), WT, and K96R mutant Sam68 vectors in response to CWP (100 nM) or ICG-001 (3 mM) 48-hr treatments. Data are expressed as relative counts versus vehicle control treatment (n = 6; *p = 0.0377, ***p % 0.0001). Error bars represent the SEM. See also Figures S6D and S6E.

Journal: Cell chemical biology

Article Title: Sam68 Allows Selective Targeting of Human Cancer Stem Cells.

doi: 10.1016/j.chembiol.2017.05.026

Figure Lengend Snippet: Figure 6. SUMOylation of Sam68 Participates in CWP Response to CSCs (A) Representative immunoﬂuorescence images of Sam68 (red) and SUMO1 (green) distribution in control and CWP-treated (100 nM, 48 hr) t-PSC (n = 8). Hoechst (blue) was used as nuclear counterstaining. Magniﬁcation: 320. (B) Dot-blot estimation of global SUMOylation levels (total SUMO1) in human PSCs versus t-PSCs, and cord blood MNCs versus primary AML samples (n = 9). Samples used: #9, #10, and #11 (Table S2). (C) Immunodetection of SUMOylated Sam68 species in vehicle, CWP, and ICG-001 treated t-PSCs (88 kDa). Puriﬁed 6xHis-Sam68 in vitro SUMOylated was used as a positive control. GAPDH performed on input lysates was used as loading control. Quantitative analysis of t-PSCs (Sam68 immunoprecipitation [IP] + western blot [WB]: n = 3) and AML cells (total Sam68 WB: n = 3) showed increased levels of SUMOylated Sam68 in response to CWP (*p = 0.0214, #p = 0.0611). See also Figures S6A–S6C. (D) Schematic illustration of wild-type (WT) and semi-SUMOylable mutant (K96R) Sam68 constructs used for overexpression experiments in t-PSCs. (E) HCI assessment of Sam68 nuclear levels from GFP-positive populations for control vector (EGFP), as well as Sam68 WT and K96R mutant transduced t-PSCs treated with CWP (100 nM) or ICG-001 (3 mM). Data are expressed as relative amounts versus control EGFP for each treatment (n = 6, **p % 0.0094). Repre- sentative micrographs for CWP versus vehicle controls are also presented. Magniﬁcation: 320. (F) Cell counts of GFP-positive populations from t-PSCs transduced with control (EGFP), WT, and K96R mutant Sam68 vectors in response to CWP (100 nM) or ICG-001 (3 mM) 48-hr treatments. Data are expressed as relative counts versus vehicle control treatment (n = 6; *p = 0.0377, ***p % 0.0001). Error bars represent the SEM. See also Figures S6D and S6E.

Article Snippet: SUMOylated recombinant Sam68 (Origene TP300263) used as positive control was in vitro modified using a SUMOylation Kit (ENZO Life Sciences).

Techniques: Control, Dot Blot, Immunodetection, In Vitro, Positive Control, Immunoprecipitation, Western Blot, Mutagenesis, Construct, Over Expression, Plasmid Preparation, Transduction

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