wi38 human diploid embryonic lung fibroblasts cells  (ATCC)

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: NaDC3-induced senescence-related phenotypes in WI38 cells. (A) Efficiency of infection with NaDC3 was detected by Western blot. The WI38/hNaDC3 cells showed significantly high level of NaDC3, but no expression was found in control groups ( P < 0.05) ( n = 6). Empty vector groups had no difference compared to control ( P > 0.05) ( n = 6). Relative NaDC3 protein band density was normalized to β-actin. * P < 0.05 versus control group. (B) Morphology observation in the WI38 cells infected with NaDC3 vector. WI38/hNaDC3, WI38/neo and WI38/con cells cultured for 37PD were observed for morphology. (C) SA-beta-gal staining in the WI38 cells infected with NaDC3 retroviral vector. WI38 cells cultured for 37PD were stained for SA-beta-gal, which showed blue precipitation in the cytoplasm in senescent cells. Magnification, 100×. (D) Analysis of SAHF formation. The cells were stained with DAPI and heterochromatin foci were shown in the senescent cells. (E) Analysis of cell cycle by flow cytometry.

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: Infection, Western Blot, Expressing, Control, Plasmid Preparation, Cell Culture, Staining, Retroviral, Flow Cytometry

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: SIRT1 inhibited Senescence-related phenotypes in WI38 cells. The SA-beta-gal staining and DAPI staining in WI38 cells were observed. The SA-beta-gal activity (A) and SAHF formation (B) were observed in the WI38 cells treated with resveratrol or niacinamide.

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: Staining, Activity Assay

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: NaDC3 inhibited SIRT1 Activity in WI38 cells. SIRT1 activity of WI38/hNaDC3 cells was inhibited dramatically, compared with those in control and WI38/neo groups ( P < 0.05). * P < 0.05 versus control group ( n = 6).

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: Activity Assay, Control

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: Elevation of SIRT1 activity reduced senescence-related phenotypes induced by NaDC3. (A) SIRT1 activity was measured using the Fluor de Lys kit in WI38/hNaDC3 cells in the presence of CR serum or resveratrol cultured for 37PD. WI38/hNaDC3 cells in the presence of CR serum or resveratrol showed significantly higher levels of SIRT1 than that in WI38/hNaDC3 group ( P < 0.05). * P < 0.05 versus WI38/hNaDC3 group ( n = 6). (B) Effect of elevating SIRT1 activity on morphology and SA-beta-gal staining of WI38/hNaDC3 cells. WI38/hNaDC3 cells cultured in the presence CR serum or resveratrol for 37 PD were stained for SA-beta-gal. The percentage of SA-beta-gal stained positive cells. For each group, 100–200 cells were counted. Comparison among groups was conducted with ANOVA. * P < 0.05 versus WI38/hNaDC3 group. Magnification, 100×. (C) SAHF formations were observe in the WI38/hNaDC3 cells treated with resveratrol or CR serum. (D) Analysis of cell cycle by flow cytometry in the WI38/hNaDC3 cells treated with resveratrol or CR serum.

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: Activity Assay, Cell Culture, Staining, Comparison, Flow Cytometry

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: Exogenous NAD + delayed cellular premature senescence induced by NaDC3. (A) NAD + /NADH ratio of WI38/hNaDC3 cells was down-regulated. But WI38/hNaDC3 + NAD cells were noted NAD + /NADH ratio was enhanced, compared with WI38/hNaDC3 cells in 37 PD ( P < 0.05). * P < 0.05 versus WI38/hNaDC3 group. (B) SIRT1 activity was measured in WI38/hNaDC3 + NAD + cells. NAD + enhanced SIRT1 activity of WI38/hNaDC3 cells, compared with WI38/hNaDC3 cells without NAD + in 37 PD ( P < 0.05). * P < 0.05 versus WI38/hNaDC3 group. The data shown is one representative experiment out of a series of five with similar results. (C) The change of morphology and the percentage of SA-beta-gal stained positive cells. For each group in 37 PD, 100–200 cells were counted. Comparison among groups was conducted with ANOVA. * P < 0.05 versus WI38/hNaDC3 group. Magnification, 100×. (D) SAHF formations were observed in the WI38/hNaDC3 cells treated with NAD + .

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: Activity Assay, Staining, Comparison

Journal: Mechanisms of Ageing and Development

Article Title: High-affinity Na + -dependent dicarboxylate cotransporter promotes cellular senescence by inhibiting SIRT1

doi: 10.1016/j.mad.2010.08.006

Figure Lengend Snippet: A proposed signaling pathway involved in NaDC3-induced aging in the WI38 cells.

Article Snippet: Human embryonic lung diploid fibroblasts WI38 and MRC-5 were obtained from ATCC.

Techniques: