trypan blue live/dead cell exclusion  (Thermo Fisher)

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Curcumin Inhibits the PPARδ-p-Akt-GLUT1 Pathway and Ameliorates the Antiproliferative Effects of Doxorubicin in MDA-MB-231 Cells

doi: 10.31557/APJCP.2024.25.3.1035

Figure Lengend Snippet: Expression of GLUT1 in Mammary Carcinoma Cells. (a) Extracts from MCF-7 and MDA-MB-231 cells were resolved with SDS-PAGE and immunoblotted with anti- GLUT1 . GAPDH was used for normalization. Densitometry analyses provide relative amount of GLUT1 normalized to GAPDH . Data are mean ± SE (n=3). *, p<0.05 (b) Total RNA was collected from MCF-7 and MDA-MB-231 cells and GLUT1 mRNA was analyzed by qRT-PCR. GAPDH was used for normalization. Data are mean ± SE (n=3).

Article Snippet: Trypan Blue Exclusion Assay MDA-MB-231 cells were transfected with control or GLUT1 siRNA overnight using Lipofectamine RNAimax and treated with 0.5 μM doxorubicin for 48 h. Cells were trypsinized, collected and counted using trypan blue from MP Biomedicals (Solon, OH, USA).

Techniques: Expressing, SDS Page, Quantitative RT-PCR

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Curcumin Inhibits the PPARδ-p-Akt-GLUT1 Pathway and Ameliorates the Antiproliferative Effects of Doxorubicin in MDA-MB-231 Cells

doi: 10.31557/APJCP.2024.25.3.1035

Figure Lengend Snippet: Inhibition of GLUT1 Improves the Efficiency of Doxorubicin-mediated Growth Suppression in MDA-MB-231. MDA-MB-231 cells were treated with 0.5 μM doxorubicin (DOX) with or without WZB117 at 6 μM (a) or 10 μM (b) for 48 h, after which growth was assessed. Data are mean ± SE of five independent experiments for panel a and four independent experiments for panel b. *: p=0.04, **: p=0.0004. (c) MDA-MB-231 cells were treated with varying concentrations of WZB117 with or without 0.5 μM DOX for 48 h. Data are mean ± SE of three independent experiments *: < p,0.05. (d) MDA-MB-231 cells were transfected with control or GLUT1 siRNA for 48 h, protein extracted, cell lysate resolved in SDS-PAGE and immunoblotted with GLUT1. Loading control: GAPDH . Transfected MDA-MB-231 cells with control or GLUT1 siRNA were treated with 0.5 μM DOX for 48 h and live cells were counted using trypan blue. Data are mean of ± SE (n=3). * p= 0.02 (versus control siRNA), ** p=0.03 (versus control siRNA + DOX), *** p=0.02 (versus GLUT1 siRNA). (e) MDA-MB-231 cells were transfected with GLUT 1 expression plasmid or control vector, treated with 0.5 μM DOX for 48 h and live cells were counted using trypan blue. Data are mean of ± SE (n=3). * p= 0.04 (versus control vector), ** p= 0.03 (versus GLUT1 OE), *** p= 0.02 (versus control vestor + Dox)

Article Snippet: Trypan Blue Exclusion Assay MDA-MB-231 cells were transfected with control or GLUT1 siRNA overnight using Lipofectamine RNAimax and treated with 0.5 μM doxorubicin for 48 h. Cells were trypsinized, collected and counted using trypan blue from MP Biomedicals (Solon, OH, USA).

Techniques: Inhibition, Transfection, SDS Page, Expressing, Plasmid Preparation

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Curcumin Inhibits the PPARδ-p-Akt-GLUT1 Pathway and Ameliorates the Antiproliferative Effects of Doxorubicin in MDA-MB-231 Cells

doi: 10.31557/APJCP.2024.25.3.1035

Figure Lengend Snippet: Downregulation of GLUT1 by Curcumin Promotes the Antiproliferative Effects of Doxorubicin. (a) MDA-MB-231 cells were treated with 20 μM and 30 μM of curcumin for 24 h, cell lysate resolved with SDS-PAGE and immunoblotted against GLUT1. (b) MDA-MB-231 cells were transfected with GLUT1 expression plasmid or control vector, treated with 0.5 μM DOX and/or 30 μM Curcumin (CUR) for 48 h and live cells were counted using trypan blue. Data are mean of ± SE (n=3). *p<0.05, **, p<0.01, ***, p<0.001

Article Snippet: Trypan Blue Exclusion Assay MDA-MB-231 cells were transfected with control or GLUT1 siRNA overnight using Lipofectamine RNAimax and treated with 0.5 μM doxorubicin for 48 h. Cells were trypsinized, collected and counted using trypan blue from MP Biomedicals (Solon, OH, USA).

Techniques: SDS Page, Transfection, Expressing, Plasmid Preparation

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Curcumin Inhibits the PPARδ-p-Akt-GLUT1 Pathway and Ameliorates the Antiproliferative Effects of Doxorubicin in MDA-MB-231 Cells

doi: 10.31557/APJCP.2024.25.3.1035

Figure Lengend Snippet: PPARδ Reverses the Downregulation of GLUT1 by Curcumin. (a) MDA-MB-231 cells transfected with control or PPARδ siRNA for 48 h were probed for PPARδ and GLUT1 . (b) Control or PPARδ expression plasmid were transfected in MDA-MB-231 cells for 48 h and probed for the protein expression of PPARδ and GLUT1 (c) MDA-MB-231 cells transfected with control or PPARδ expression plasmid overnight were treated with or without 30 μM of curcumin 24 h post-transfection, and probed for GLUT1 protein. Data depicts a representation of a Western blot from three independent experiments (a-c)

Article Snippet: Trypan Blue Exclusion Assay MDA-MB-231 cells were transfected with control or GLUT1 siRNA overnight using Lipofectamine RNAimax and treated with 0.5 μM doxorubicin for 48 h. Cells were trypsinized, collected and counted using trypan blue from MP Biomedicals (Solon, OH, USA).

Techniques: Transfection, Expressing, Plasmid Preparation, Western Blot

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Curcumin Inhibits the PPARδ-p-Akt-GLUT1 Pathway and Ameliorates the Antiproliferative Effects of Doxorubicin in MDA-MB-231 Cells

doi: 10.31557/APJCP.2024.25.3.1035

Figure Lengend Snippet: PPARδ and Akt are Involved in the Regulation of Curcumin-mediated Downregulation of GLUT1 (a) MDA-MB-231 cells were treated with 30 μM of curcumin or control for 24 h to probe for p-Akt and Akt. MDA-MB-231 cells were transfected with PPARδ (b) or Akt (c) expression plasmids, treated with 30 μM of curcumin and probed for p-Akt (b) or GLUT1 (c). Loading control: GAPDH. Data depicts a representation of a Western blot from three independent experiments (a-c) (d) A model outlining the effects of curcumin on the interplay between PPARδ, Akt and GLUT1

Article Snippet: Trypan Blue Exclusion Assay MDA-MB-231 cells were transfected with control or GLUT1 siRNA overnight using Lipofectamine RNAimax and treated with 0.5 μM doxorubicin for 48 h. Cells were trypsinized, collected and counted using trypan blue from MP Biomedicals (Solon, OH, USA).

Techniques: Transfection, Expressing, Western Blot