Journal: Journal of Cellular and Molecular Medicine

Article Title: Overexpression of sortilin is associated with 5‐FU resistance and poor prognosis in colorectal cancer

doi: 10.1111/jcmm.15752

Figure Lengend Snippet: Reponses to 5‐FU short‐term treatment (8 µmol/L, 72 h) of WiDr and SW620: metabolic activity, cell death and sortilin expression. WiDr and SW620 were treated with 5‐FU for 72 h. (A, Increased 5‐FU concentrations were tested for metabolic activity analysis using MTT assay (Promega, France). Untreated cells were used as control (100%). B, Cell death induction after 5‐FU treatment (8 µmol/L, 72 h) was determined by flow cytometry with PI/AnnexinV‐FITC. C, Sortlin protein expression was analysed by Western blotting from whole‐cell lysates of 5‐FU‐treated WiDr and SW620 (8 µmol/L, 72 h) and ‐untreated cells (CTL). Actin was used as loading control. Histograms are the means from at least three independent experiments. Significant P‐values are indicated in the graphs * P < .05, ** P < .01, *** P < .001

Article Snippet: Seventy‐two hours later, metabolic activity was determined with the thiazolyl blue tetrazolium blue (MTT) assay (CellTiter 96 Aqueous One solution Cell Proliferation Assay, Promega, Charbonnières‐les‐Bains, France) following supplier instructions.

Techniques: Activity Assay, Expressing, MTT Assay, Control, Flow Cytometry, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Overexpression of sortilin is associated with 5‐FU resistance and poor prognosis in colorectal cancer

doi: 10.1111/jcmm.15752

Figure Lengend Snippet: Reponses to 5‐FU long‐term treatment (8 µmol/L, one month) of WiDr, SW620 and primary cultures: cell death, insensitivity to 5‐FU increasing concentrations and sortilin expression. After one month of 5‐FU treatment, A, cells were analysed by flow cytometry for detection of living and dead cells (PI/Annexin‐FITC) as described before. Histograms are means from at least three independent experiments. B, Metabolic activity of 5‐FU‐resistant cells was determined using MTT assay (Promega, France). Untreated cells (maintained in culture during one month) were also analysed and used as control (CTL). Significant P‐values are indicated in the graphs: * P < .05; ** P < .01; *** P < .001. Sortlin protein expression was analysed by Western blotting from whole‐cell lysates of 5‐FU‐resistant (5‐FU) and ‐untreated cells (CTL) in WiDr and SW620 C, and in primary cultures D,. Actin was used as loading control. Histograms are the means from at least three independent experiments. Significant P‐values are indicated in the graphs * P < .05, ** P < .01, *** P < .001. E, Sortilin transcriptomic expression of 5‐FU‐resistant cells (one month) and ‐untreated cells (CTL) was analysed by RT‐q‐PCR. Sortilin mRNA expression ( SORT1 ) upon 5‐FU was normalized to both HPRT transcriptomic expression and that of untreated cells. Histograms are the means from at least three independent experiments. Significant P‐values are indicated in the graphs * P < .05, ** P < .01, *** P < .001

Article Snippet: Seventy‐two hours later, metabolic activity was determined with the thiazolyl blue tetrazolium blue (MTT) assay (CellTiter 96 Aqueous One solution Cell Proliferation Assay, Promega, Charbonnières‐les‐Bains, France) following supplier instructions.

Techniques: Expressing, Flow Cytometry, Activity Assay, MTT Assay, Control, Western Blot