Display Settings:

Items per page
We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 9

1.
Figure 3.

Figure 3. From: Smurf2 up-regulation activates telomere-dependent senescence.

Lack of DNA damage response in Smurf2-induced senescent cells. BJ cells of replicative senescence, Smurf2-induced senescence, and GFP expressing control, as well as early passage cells 1 h after being treated with 10 Gy of ionizing radiation (IR), were stained with anti-phospho-histone H2A.X (Ser 139) antibody and DAPI.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
2.
Figure 5.

Figure 5. From: Smurf2 up-regulation activates telomere-dependent senescence.

Elevation of Smurf2 reversed cellular immortalization by hTERT. GFP, Smurf2, and C716A were adventitiously expressed in hTERT-immortalized human fibroblast cell lines (BJ-hTERT and WS1-hTERT) and selected for puromycin resistance. (A) Western blot analysis of Smurf2 protein expression. (B) Proliferative potential of these cells was monitored as described in Materials and Methods. (C) Crystal violet and pH 6 β-galactosidase activity staining.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
3.
Figure 8.

Figure 8. From: Smurf2 up-regulation activates telomere-dependent senescence.

Absence of p21 does not abrogate the effect of Smurf2 induction. Early passage p21+/+ LF1 fibroblasts, p21-/- fibroblasts, and their hTERT-immortalized derivatives were infected with retrovirus expressing control GFP, wild-type Smurf2, or ligase mutant C716A, and selected with puromycin. (A) Western blot analysis of protein expression in these cells. (B) Growth curves of hTERT-immortalized LF1, p21-null cells after adventitious expression of GFP, Smurf2, and C716A. (C) Crystal violet and β-galactosidase activity at pH 6 staining in hTERT-immortalized LF1, p21-null cells after adventitious expression of GFP, Smurf2, and C716A.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
4.
Figure 9.

Figure 9. From: Smurf2 up-regulation activates telomere-dependent senescence.

Expression of HPV16 E6 and E7 abrogates Smurf2-induced senescence. Early passage BJ fibroblasts were infected with lentivirus expressing HPV16 E6, E7 alone, or E6 and E7 together. In these E6/E7-expressing populations, Smurf2 as well as GFP control were ectopically expressed using retrovirus. (A) Western blot analysis of protein expression. (B) Growth curves of BJ-E6, BJ-E7, and BJ-E6-E7 populations after adventitious expression of GFP, Smurf2, and C716A. (C) Crystal violet and β-galactosidase activity at pH 6 staining of BJ-E6, BJ-E7, and BJ-E6-E7 populations after adventitious expression of GFP, Smurf2, and C716A.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
5.
Figure 7.

Figure 7. From: Smurf2 up-regulation activates telomere-dependent senescence.

Concurrent elevation of p21 and cyclin D1 during Smurf2-induced senescence as well as during replicative senescence. (A) Western blot analysis of various proteins in fibroblasts that are early passage proliferating (EP), quiescent (Q) by contact inhibition, or replicative senescent (RS) by serial passage. (B) Western blot analysis of various proteins in early passage human fibroblasts (BJ, WS1, and WI38) infected with retrovirus expressing control GFP, wild-type Smurf2, or ligase mutant C716A. (C) Protein expression in hTERT-immortalized fibroblasts after adventitiously expressing wild-type Smurf2, ligase mutant C716A, and control GFP.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
6.
Figure 6.

Figure 6. From: Smurf2 up-regulation activates telomere-dependent senescence.

Domains of Smurf2 that are necessary for its senescence-inducing function. Different portions of Smurf2, including the C2 domain (amino acids 1-147), three WW domains (amino acids 149-344), the HECT domain (amino acids 345-748), N-ter (amino acids 1-344), ΔC2 (amino acids 149-748), and ΔWW1 (amino acids 242-748), were cloned into pBabe-IRES-Puro using cDNA generated by PCR, and ectopically expressed in two early passage fibroblast lines (WS1 and WI38) and a human fibrosarcoma cell line HT1080. After selection with puromycin, plates were stained with crystal violet.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
7.
Figure 4.

Figure 4. From: Smurf2 up-regulation activates telomere-dependent senescence.

Microarray analysis of gene expression alterations in Smurf2-induced senescent cells. Transcripts from senescent cells induced by Smurf2 or its ligase mutant C716A were labeled with Cy5-dUTP (red), and hybridized onto microarrays with Cy3-dUTP (green)-labeled transcripts from GFP-expressing control cells. Data for replicative senescence in fibroblasts were retrieved from a previously published study (Zhang et al. 2003). In that data set, transcripts from replicative senescence (labeled with Cy5-dUTP) were compared with those from early passage proliferating cells (labeled with Cy3-dUTP). Data were analyzed using GABRIEL as described in Materials and Methods. (A) Transcriptional alterations in common between Smurf2-induced senescence and telomere-mediated replicative senescence. (B) Transcriptional alterations occurring only during replicative senescence but not during Smurf2-induced senescence. (C) Transcriptional alterations induced by Smurf2 elevation only.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
8.
Figure 2.

Figure 2. From: Smurf2 up-regulation activates telomere-dependent senescence.

Adventitious expression of Smurf2-induced senescence in early passage human fibroblasts (BJ, WS1, and WI38). (A) Western blot analysis of Smurf2 protein in early passage cells infected with retrovirus expressing GFP, Smurf2, or ligase mutant C716A and selected for puromycin resistance. (B) Proliferative potentials of early passage cells infected with retrovirus containing GFP, Smurf2, or ligase mutant C716A. After selection for puromycin resistance, infected cells were plated at low density and cell numbers were counted daily. A represented growth curve for each cell line was shown. (C) Crystal violet staining of early passage cells after infection with retrovirus containing GFP, Smurf2, or ligase mutant C716A. (D) Staining for β-galactosidase activity at pH 6 of early passage cells after infection with retrovirus containing GFP, Smurf2, or ligase mutant C716A.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.
9.
Figure 1.

Figure 1. From: Smurf2 up-regulation activates telomere-dependent senescence.

Increased expression of Smurf2 during replicative senescence in human fibroblasts. (A) Northern analysis of Smurf2 expression in cells that are early passage proliferating (EP), quiescent (Q) by contact inhibition, and replicative senescent (RS) by serial passage. The probe used was the cDNA of Smurf2 and GAPDH as loading control. Two fibroblast lines (WS1 and WI38) and two HMEC lines (48R and 184) were analyzed. (B) Western blot analysis of Smurf2 protein level in human fibroblasts that are early passage proliferating (EP), quiescent (Q), or senescent (RS). Three fibroblast lines (BJ, WS1, and WI38) were used. α-Tubulin was used as loading control. (C) Expression of hTERT and consequent immortalization prevents the induction of Smurf2. Western blot analysis of the Smurf2 protein level in human fibroblasts (BJ and WS1) of early passage (EP) and hTERT-immortalized (T) cells. (D) Expression of Smurf2 is not induced by oxidative stress. Western blot analysis in early passage fibroblasts treated with a sublethal dose of H2O2 (250 μM). (E) DNA damage response activated by ionizing radiation does not induce Smurf2. Early passage BJ and WS1 were irradiated with different doses, and cells were analyzed 1, 24, 48, 96, or 168 h after irradiation using Western blotting. (F) Expression of Smurf2 protein is not elevated by the expression of the dominant-negative allele of TRF2 (TRF2ΔBΔM). Early passage BJ and WS1 fibroblasts were infected with retrovirus to express Myc-tagged TRF2 or TRF2ΔBΔM, or GFP control, selected with puromycin resistance for a week, and analyzed using Western blotting.

Hong Zhang, et al. Genes Dev. 2004 December 15;18(24):3028-3040.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk