flow cytometry hct116 cells  (Novus Biologicals)

Journal: Non-coding RNA Research

Article Title: Telomerase RNA component lncRNA as potential diagnostic biomarker promotes CRC cellular migration and apoptosis evasion via modulation of β-catenin protein level

doi: 10.1016/j.ncrna.2023.03.004

Figure Lengend Snippet: LncRNAs profiling in CRC cell lines. Volcano plot of the pairwise comparison of lncRNAs expression in (A) HCT116 cell line and (B) SW620 cell line versus control cell line. Overexpressed lncRNAs are shown to the right of the plot (green) and were only selected if they passed the thresholds of false discovery rate (FDR) > 2 (horizontal blue line) and log2 fold change >1 (right vertical line). Accordingly, down-regulated lncRNAs are shown to the left of the plot (yellow) and were only selected if they passed the thresholds of FDR >2 (horizontal blue line) and log2 fold change <-1 (left vertical line). (C) Clustering graph of the lncRNA different expression level between the CRC cell lines. Significant from control at p < 0.05 and analyzed by two-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Flow cytometry HCT116 cells transfected with si-TERC and si-NC were harvested 48 h later for cell cycle analysis Phycoerythrin was used to stain cells with the PE-Texas Red™ Antibody Labeling Kit (Novus Biologicals™ 7670030).

Techniques: Comparison, Expressing, Control

Journal: Non-coding RNA Research

Article Title: Telomerase RNA component lncRNA as potential diagnostic biomarker promotes CRC cellular migration and apoptosis evasion via modulation of β-catenin protein level

doi: 10.1016/j.ncrna.2023.03.004

Figure Lengend Snippet: The wound healing was used for migration assay. (A) Images taken to HCT116 control cells, si-NC transfected cells, and si-TERC transfected cells at different time intervals. (B) The mean ± SD of wound width was significantly larger in si-TERC transfected cells than control and si-NC after 24 h. (C) The mean ± SD of migration rate was significantly decreased in si-TERC transfected cells than control and si-NC after 48 h. (D) The mean ± SD of wound area was significantly larger in si-TERC transfected cells than control and si-NC after 24 h. (E) The mean ± SD of wound closure was significantly decreased in si-TERC transfected cells than control and si-NC after 48 h *Significantly different in compare to control and si-NC at p < 0.05. This data was assessed by two-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Flow cytometry HCT116 cells transfected with si-TERC and si-NC were harvested 48 h later for cell cycle analysis Phycoerythrin was used to stain cells with the PE-Texas Red™ Antibody Labeling Kit (Novus Biologicals™ 7670030).

Techniques: Migration, Control, Transfection

Journal: Non-coding RNA Research

Article Title: Telomerase RNA component lncRNA as potential diagnostic biomarker promotes CRC cellular migration and apoptosis evasion via modulation of β-catenin protein level

doi: 10.1016/j.ncrna.2023.03.004

Figure Lengend Snippet: (A) Cell cycle analysis of HCT116 cell lines. (B) The mean ± SD of number of HCT116 cells were significantly increased in the G0/G1 phase after lncRNA TERC knockdown in comparing to control and si-N. In addition, the mean ± SD of number of HCT116 cells were significantly decreased in the S phase in comparing to control and si-NC. *Significantly different in compare to control and si-NC at p < 0.05. This data was assessed using two-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Flow cytometry HCT116 cells transfected with si-TERC and si-NC were harvested 48 h later for cell cycle analysis Phycoerythrin was used to stain cells with the PE-Texas Red™ Antibody Labeling Kit (Novus Biologicals™ 7670030).

Techniques: Cell Cycle Assay, Knockdown, Control

Journal: Non-coding RNA Research

Article Title: Telomerase RNA component lncRNA as potential diagnostic biomarker promotes CRC cellular migration and apoptosis evasion via modulation of β-catenin protein level

doi: 10.1016/j.ncrna.2023.03.004

Figure Lengend Snippet: (A) Flow cytometric analysis for cell apoptosis. (B) The mean ± SD of percentage of HCT116 apoptotic cells were significantly increased after lncRNA TERC knockdown in comparing to control and si-NC. *Significantly different in compare to control and si-NC at p < 0.001. This data was assessed using two-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Flow cytometry HCT116 cells transfected with si-TERC and si-NC were harvested 48 h later for cell cycle analysis Phycoerythrin was used to stain cells with the PE-Texas Red™ Antibody Labeling Kit (Novus Biologicals™ 7670030).

Techniques: Knockdown, Control

Journal: Non-coding RNA Research

Article Title: Telomerase RNA component lncRNA as potential diagnostic biomarker promotes CRC cellular migration and apoptosis evasion via modulation of β-catenin protein level

doi: 10.1016/j.ncrna.2023.03.004

Figure Lengend Snippet: β-catenin expression pattern in transfected HCT 116 cell line (A) The mean ± SD expression fold change of β-catenin mRNA in HCT 116 cell lines (B) Normalized level in TERC knockdown HCT116 cells and control cells. *Significantly different in compare to control and si-NC at p < 0.001. This data was assessed using two-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: Flow cytometry HCT116 cells transfected with si-TERC and si-NC were harvested 48 h later for cell cycle analysis Phycoerythrin was used to stain cells with the PE-Texas Red™ Antibody Labeling Kit (Novus Biologicals™ 7670030).

Techniques: Expressing, Transfection, Knockdown, Control

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: ( A ) Expression of SMAR1 and β-catenin in various CRC cell lines. ( B ) Confocal staining of colon tissue sections co-stained with SMAR1 and β-catenin antibodies. Arrow shows the basal portion of the colon crypt. The scale bar used in the confocal experiment represents 30 μm. ( C ) Expression of SMAR1 and β-catenin levels in mouse colon tissues (polyp vs normal adjacent tissue). ( D and E ) Expression of SMAR1 and β-catenin upon stimulating HCT116 cells with Wnt3a CM or rh Wnt3a ligand (200 ng/mL). ( F ) Confocal staining of SMAR1 after Wnt3a CM stimulation in HCT116 cells. The scale bar used in the confocal experiment represents 20 μm. ( G ) SMAR1 expressions after treating HCT116 cells with both Wnt3a CM and 10 μM MG132 drug. ( H ) SMAR1 expression in FLAG-SMAR1, D1, D2 and D3 expressing cells after Wnt3a CM stimulation.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Expressing, Staining

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: ( A ) β-catenin expression in HCT116 cells after knockdown with sh-SMAR1. The triangle indicates increased concentration of SMAR1 plasmids of 1 and 3 μg. ( B ) Real-Time PCR experiment showing β-catenin mRNA levels in SMAR1 knockdown HCT116 cells. 18S rRNA was used for normalization. Results are shown as mean ± SD ( n = 3). The p value was determined by student's t -test. The triangle indicates increased concentration of SMAR1 plasmids of 2 and 4 μg. ( C ) β-catenin expressions in HCT116 cells after GFP-SMAR1 overexpression. The triangle indicates increased concentration of SMAR1 plasmids of 1, 2, 3 and 4 μg. ( D ) Real Time-PCR experiment showing β-catenin mRNA levels in SMAR1 overexpressed HCT116 cells. 18s rRNA was used for normalization. Results are shown as mean ± SD ( n = 3). The p value was determined by student's t -test. The triangle indicates increased concentration of SMAR1 plasmids of 1, 2.5 and 4 μg. ( E ) Luciferase promoter activities of Super 8X TOPFlash/FOPFlash (mean ± SD, n = 3) after SMAR1 overexpression and knockdown in HCT116 cells. ( F ) Expression of β-catenin in HCT116 cells after transfection with GFP-SMAR1 and simultaneous stimulation with Wnt3a CM.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Expressing, Knockdown, Concentration Assay, Real-time Polymerase Chain Reaction, Over Expression, Luciferase, Transfection

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: FACS analysis of pEGFP1-β-catenin GFP expression ( n = 3, SD) after; ( A ) Co-transfection with FLAG-vector or FLAG-SMAR1, and ( B ) Treatment with 200 ng/mL rh Wnt3a ligand. ( C ) ChIP showing occupancy of SMAR1, HDAC5 and H3K9 Ac (mean ± SD, n = 3) after Wnt3a CM stimulation. ( D ) ChIP showing occupancy of SMAR1, HDAC5 and H3K9 Ac (mean ± SD, n = 3) after SMAR1 overexpression or knockdown. ( E ) ChIP showing occupancy of HDAC5 (mean ± SD, n = 3) after si-HDAC5 knockdown in HCT116 cells. ( F ) Expression of β-catenin after knockdown with 2μg si-HDAC5 plasmid. ( G ) Immunoprecipitation of SMAR1 with HDAC5. ( H ) Immunoprecipitation of HDAC5 with various truncations of SMAR1. ( I ) Immunoprecipitation of HDAC5 with SMAR1 after Wnt3a CM stimulation.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Expressing, Cotransfection, Plasmid Preparation, Over Expression, Knockdown, Immunoprecipitation

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: ( A ) Cell invasion in SW480 cells after overexpression with GFP-SMAR1 construct. ( B ) Cell migration assay in GFP-SMAR1 overexpressed SW480 cells. ( C ) Tumors generated in NOD-SCID mice ( n = 10) using various stable HCT116 cells for SMAR1. ( D and E ) Graphs showing volume and weight of tumors generated in NOD-SCID mice. ( F ) Kaplan Meier survival probability curve plotted with respect to SMAR1 expression in CRC patients. The survival curve was generated using Smith tumor colon database.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Over Expression, Construct, Cell Migration Assay, Generated, Expressing

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: ( A ) Expression of SMAR1 in HCT116 cells treated with various peptides (5 μg/mL) for 48 hrs. ( B ) Confocal staining for SMAR1 after AT-01C or AT-01D (10 μg/mL) treatment. The scale bar used in the confocal experiment represents 10 μm. β-catenin expression after treatment of HCT116 cells with increasing concentration of; ( C ) AT-01C and ( D ) AT-01D peptides. SMAR1 expression after stimulation of HCT116 cells with Wnt3a CM and simultaneously treated with ( E ) AT-01C, and ( F ) AT-01D. ( G ) Luciferase promoter activities of Super 8X TOPFlash/FOPFlash (mean ± SD, n = 3) after treatment with AT-01C or AT-01D.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Expressing, Staining, Concentration Assay, Luciferase

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: Isothermal Titration Calorimetry (ITC) showing the interaction of SMAR1 with the peptides, ( A ) AT-01C and ( B ) AT-01D. In silico AutoDock interactions of SMAR1 with: ( C ) AT-01C and ( D ) AT-01D peptide. ( E ) SMAR1 expression after treating HCT116 cells with single peptide or combination of AT-01C and AT-01D.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Isothermal Titration Calorimetry, In Silico, Expressing

Journal: Oncotarget

Article Title: SMAR1 inhibits Wnt/β-catenin signaling and prevents colorectal cancer progression

doi: 10.18632/oncotarget.25093

Figure Lengend Snippet: ( A ) Cell migration assay in SW480 cells treated with AT-01C or AT-01D for 12 hours. ( B ) Graphical representation of SW480 cells migrated (mean ± SD, distance in triplicates arbitrarily taken at three different points). ( C ) Cell invasion assays in SW480 cells after treatment with AT-01C or AT-01D (10 μg/mL) for 48 hrs. ( D ) Clonogenic assays in HCT116 cells after treatment with AT-01C or AT-01D (10 μg/mL), and ( E ) Its graphical representation ( n = 3, SD). ( F ) Tumors generated using 1 × 10 6 HCT116 cells in NOD-SCID mice after administration with AT-01C (25 mg/kg body weight). ( G and H ) Graph representing the weight and volume of the mice tumors administered with AT-01C. ( I ) Tumors generated using 1 × 10 6 HCT116 cells in NOD-SCID mice after administration with AT-01D (25 mg/kg body weight). ( J and K ) Graph representing the weight and volume of the mice tumors administered with AT-01D.

Article Snippet: For flow cytometry analysis HCT116 cells were trypsinized using Trypsin-EDTA (Gibco) and washed thrice with PBS buffer.

Techniques: Cell Migration Assay, Generated