Journal: bioRxiv

Article Title: A genetic screen to identify deubiquitinases as regulators of IRF7

doi: 10.1101/2025.09.09.675186

Figure Lengend Snippet: (A) RAW-Luci macrophages were either infected with SeV, or transfected with poly(I:C) (pIC) using Lipofectamine (LF), or treated with pIC alone. Otud5 mRNA levels were quantified by qRT-PCR. (B–G) RAW264.7 cells were transfected with NT, Otud5 (O5), or Usp2 (U2) siRNAs, infected with SeV for 8 hours, and mRNA levels of Otud5, Usp2, Ifna, Ifnb1, Ifit1, and Ifit3 were analyzed by qRT-PCR. (H–J) HEK-KO.IRF7 cells were transfected with NT, OTUD5 (O5), or USP2 (U2) siRNAs and infected with SeV. USP2 , OTUD5 , and IFNA mRNA levels were measured by qRT-PCR. The data represent mean ± SEM (A-G, J), * p<0.05, ** p<0.001, *** p<0.001, **** p<0.0001.

Article Snippet: Plasmids included HA-Ub-K27O (Addgene #22903), HA-Ub-K63O (Addgene #17606), OTUD5 (Sino Biologicals #HG22452-CF), and USP2 (Sino Biologicals #HG12713-UT).

Techniques: Infection, Transfection, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A genetic screen to identify deubiquitinases as regulators of IRF7

doi: 10.1101/2025.09.09.675186

Figure Lengend Snippet: (A) USP2:IRF7 interaction in HEK-KO.IRF7 cells was assessed by co-immunoprecipitation at the indicated times post-SeV infection. (B–C) Proximity ligation assay (PLA) was performed in WT primary BMDMs using anti-USP2 and anti-IRF7 antibodies in mock or SeV-infected (8 hpi) cells. Red dots represent interaction signals, quantified in (C). (D) OTUD5:IRF7 interaction in HEK-KO.IRF7 cells was analyzed by co-immunoprecipitation following SeV infection. (E–F) PLA was conducted in WT BMDMs using anti-OTUD5 and anti-IRF7 antibodies in mock or SeV-infected cells. Interaction signals were quantified in (F).

Article Snippet: Plasmids included HA-Ub-K27O (Addgene #22903), HA-Ub-K63O (Addgene #17606), OTUD5 (Sino Biologicals #HG22452-CF), and USP2 (Sino Biologicals #HG12713-UT).

Techniques: Immunoprecipitation, Infection, Proximity Ligation Assay

Journal: bioRxiv

Article Title: A genetic screen to identify deubiquitinases as regulators of IRF7

doi: 10.1101/2025.09.09.675186

Figure Lengend Snippet: (A) HEK-KO.IRF7 cells were transfected with Flag.OTUD5 or USP2 plasmids, and the cell lysates were analyzed by immunoblot, as indicated. (B–D) HEK-KO.IRF7 cells were transfected with HA-tagged Ub-K63O, Ub-K27O, or Ub-K33O plasmids, in the absence or the presence of OTUD5 or USP2, as indicated, and infected with SeV. The cell lysates were immunoprecipitated with anti-V5 antibody and immunoblotted with anti-HA, as indicated. (E–G) Full-length and truncated IRF7 mutants (E) were co-transfected with Ub-K63O or Ub-K27O, followed by SeV infection. Ub-IRF7 levels were measured by IP and immunoblot (F) and quantified using ImageJ (G). (H) Schematic of IRF7 protein domains and putative ubiquitin linkage sites (K63 and K27); DBD, DNA-binding domain. (I) Cells were transfected with HA.Ub-K27O and infected with SeV. Ub-IRF7 was analyzed as in (A). (J) IRF7-WT or KR mutants (K1: K327R, K2: K329R, K3: K327/329RR) were co-transfected with Ub-K27O, infected with SeV, and analyzed for Ub-IRF7. The lower panel shows the expression of the IRF7 mutants. EV, empty vector; Ub-IRF7, ubiquitinated IRF7.

Article Snippet: Plasmids included HA-Ub-K27O (Addgene #22903), HA-Ub-K63O (Addgene #17606), OTUD5 (Sino Biologicals #HG22452-CF), and USP2 (Sino Biologicals #HG12713-UT).

Techniques: Transfection, Western Blot, Infection, Immunoprecipitation, Ubiquitin Proteomics, Binding Assay, Expressing, Plasmid Preparation

Journal: Diabetes

Article Title: Deubiquitinating Enzyme USP2 Alleviates Muscle Atrophy by Stabilizing PPAR-γ

doi: 10.2337/db24-0375

Figure Lengend Snippet: Two lysine residues of PPAR-γ (184 and 185) are targeted for deubiquitination by USP2. A : The hot map of RNA-sequencing of Gas muscle of WT and USP2KO mice ( n = 5). B : Total cell lysates from the Gas muscle of mice subjected to Co-IP with anti-USP2 antibody and Western blots using indicated antibodies. C2C12 cells were transfected with Ad HA-Ppar-γ and/or Ad-Flag Usp2 WT, as indicated. Total cells lysates were subjected to Co-IP with anti-Flag antibody; Western blotting results used the indicated antibodies. C : Human skeletal muscle cells (HSkMCs) transfected with Ad-Flag USP2 WT (Flag-WT) or Ad-Flag USP2 C276A. Total cell lysates were subjected to Co-IP with anti-PPAR-γ antibody; Western blots using indicated antibodies, and the charts report the quantitative result. D : WT or USP2 knockdown HSkMCs transfected with Ad-Flag PPAR-γ WT, K184R, K185R, K268R, K293R, or K462R as indicated. The total cell lysates were prepared; Western blotting used the indicated antibodies; chart reports the quantitative result. * P < 0.05, ** P < 0.01, *** P < 0.001, by unpaired Student t test. E : WT or USP2 knockdown HSkMCs transfected with Ad-Flag PPAR-γ WT and DKR (both K184R and K185R) as indicated. Total cell lysates subjected to Co-IP with anti-Flag antibody; Western blots using indicated antibodies and charts of the quantitative results are shown. F : USP2–PPAR-γ docking with the HDOCK server. High magnification of boxed areas is presented on the right in each row. Arrow indicates PPAR-γ protein K184 and K185 site. Data are expressed as mean ± SD. B , C , and E : * P < 0.05, ** P < 0.01, *** P < 0.001, by one-way ANOVA with Bonferroni correction.

Article Snippet: Usp2 flox/flox mice (C57BL/6JCya- Usp2 em1flox/Cya; strain no. S-CKO-11600) were purchased from Cyagen Biosciences (Guangzhou, Guangdong, China).

Techniques: RNA Sequencing, Co-Immunoprecipitation Assay, Western Blot, Transfection, Knockdown

Journal: Diabetes

Article Title: Deubiquitinating Enzyme USP2 Alleviates Muscle Atrophy by Stabilizing PPAR-γ

doi: 10.2337/db24-0375

Figure Lengend Snippet: USP2 improves insulin resistance in skeletal muscle. A – C : Immunoblot analysis of PPAR-γ, GLUT4, IRS1, and tubulin in Gas from mice as indicated. The chart presents the levels of the indicated protein normalized to tubulin ( n = 6). D : Immunoblot analysis of PPAR-γ, USP2, GLUT4, IRS1, and tubulin in C2C12 myotubes transfected with Ctrl or Usp2 siRNA (si Usp2 ) in the presence or absence of TNF-α at 20 ng/mL for 24 h. The chart in the middle presents the levels of the indicated protein normalized to tubulin ( n = 3). The chart at the right reports the qPCR analysis of Atrogin1, MUSA1 , and F-box protein 31 ( Fbxo31 ) in C2C12 myotubes transfected with Ctrl or Usp2 siRNA and treated with or without TNF-α at 20 ng/mL for 24 h ( n = 5). E : MYHC immunofluorescence of C2C12 myotubes transfected with Ctrl or Usp2 siRNA and treated with or without TNF-α at 20 ng/mL for 24 h. The chart presents the levels of fusion index and myotube diameter ( n = 5). F : Immunoblot analysis of PPAR-γ, USP2, GLUT4, IRS1, and tubulin in C2C12 myotubes infected with adenovirus expressing Usp2 (Ad Usp2 ) or green fluorescent protein (Ctrl). Myotubes were cultured for 24 h in the presence or absence of TNF-α at 20 ng/mL. The middle chart presents the levels of the indicated protein normalized to tubulin ( n = 3). The chart on the right presents results of the qPCR analysis of Atrogin1, MUSA1 , and Fbxo31 in C2C12 myotubes infected with Ad Usp2 or Ctrl and treated with or without TNF-α at 20 ng/mL for 24 h ( n = 5). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, by one-way ANOVA with Bonferroni correction. Vehi, vehicle.

Article Snippet: Usp2 flox/flox mice (C57BL/6JCya- Usp2 em1flox/Cya; strain no. S-CKO-11600) were purchased from Cyagen Biosciences (Guangzhou, Guangdong, China).

Techniques: Western Blot, Transfection, Immunofluorescence, Infection, Expressing, Cell Culture

Journal: Diabetes

Article Title: Deubiquitinating Enzyme USP2 Alleviates Muscle Atrophy by Stabilizing PPAR-γ

doi: 10.2337/db24-0375

Figure Lengend Snippet: USP2-regulated insulin signaling depends on PPAR-γ in vitro. A : Immunoblot analysis of USP2, PPAR-γ, IRS1, GLUT4, and tubulin in C2C12 myotubes transfected with Ctrl or Pparγ siRNA (si Pparγ ) in the presence or absence of TNF-α at 20 ng/mL for 24 h. Charts present the levels of the indicated protein normalized to tubulin ( n = 3). B : qPCR analysis of Ccng2 , Cdkn1b , Rbl2 , Bnip3, Atrogin1, MUSA1 , and F-box protein 31 ( Fbxo31 ) in C2C12 myotubes ( n = 5). C : MYHC immunofluorescence of C2C12 myotubes transfected with si Usp2 and/or si Pparγ in the presence or absence of TNF-α at 20 ng/mL for 24 h. Charts present the levels of fusion index ( n = 5). D : Immunoblots analysis of USP2, PPAR-γ, IRS1, GLUT4, and tubulin in C2C12 myotubes infected with adenovirus expressing Usp2 (Ad Usp2 ) and/or si Pparγ as indicated. Myotubes were cultured for 24 h in the presence or absence of DEX at 50 μmol/L. Charts present the levels of the indicated protein normalized to tubulin ( n = 3). E : qPCR analysis of Ccng2 , Cdkn1b , Rbl2 , Bnip3, Atrogin1, MUSA1 , and Fbxo31 in C2C12 myotubes infected with Ad Usp2 and/or si Pparγ and treated with or without DEX at 50 μmol/L for 24 h ( n = 5). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, by one-way ANOVA with Bonferroni correction.

Article Snippet: Usp2 flox/flox mice (C57BL/6JCya- Usp2 em1flox/Cya; strain no. S-CKO-11600) were purchased from Cyagen Biosciences (Guangzhou, Guangdong, China).

Techniques: In Vitro, Western Blot, Transfection, Immunofluorescence, Infection, Expressing, Cell Culture

Journal: Diabetes

Article Title: Deubiquitinating Enzyme USP2 Alleviates Muscle Atrophy by Stabilizing PPAR-γ

doi: 10.2337/db24-0375

Figure Lengend Snippet: PPAR-γ-inhibition abolished the effects of USP2 KO on aggravating insulin resistant and alleviating muscle fiber atrophy. We established a DM-induced muscle atrophy model in WT mice transfected with AAV-sh Pparγ and/or AAV- Usp2 , as indicated. n = 6. A : Western blot analysis of USP2, PPARγ, MURF-1, MYHC, and tubulin in the Gas muscle of mice. Charts present the quantification results. B : The FBG and FBI of mice. C : The ratio of Gas muscle and TA muscle weight to body weight. D : Grip strength test and exhaustive running distance results. E : Representative images of myofiber cross-sections obtained through hematoxylin-eosin (H-E) staining (scale bar = 100 μmol/L) (left) and cross-sectional area (CSA) of Gas muscles from mice (right). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, by one-way ANOVA with Bonferroni correction. Vehi, vehicle.

Article Snippet: Usp2 flox/flox mice (C57BL/6JCya- Usp2 em1flox/Cya; strain no. S-CKO-11600) were purchased from Cyagen Biosciences (Guangzhou, Guangdong, China).

Techniques: Inhibition, Transfection, Western Blot, Staining, Muscles

Journal: Autophagy

Article Title: Phospho-ubiquitin-PARK2 complex as a marker for mitophagy defects

doi: 10.1080/15548627.2016.1254852

Figure Lengend Snippet: PARK2 forms a phospho-ubiquitin-rich complex with VDAC. (A) FLAG-PARK2 and control cells were differentially labeled using SILAC. Equal amounts of mitochondria were mixed, solubilized and subjected to anti-FLAG immunoprecipitation, followed by native elution. Complexes from 2 independent experiments (with label switch) were resolved using BN-PAGE. Gel lanes were cut into equal slices, within the range of the 500-kDa complex, and each slice was analyzed by LC-MS. Shown are relative protein abundance ratios (FLAG-PARK2:control). (B) Solubilized mitochondria from FLAG-PARK2 cells treated with CCCP (3 h) were analyzed by 2D BN/SDS-PAGE, followed by western blotting. Asterisk indicates likely ubiquitinated forms of VDAC. (C) Anti-FLAG immunoprecipitations using mitochondria solubilized from FLAG-PARK2-expressing cells, treated with 10 μM CCCP, or with DMSO as a control, for 3 h. Solubilized mitochondria from COX6A1-FLAG-expressing cells were used as a specificity control.26 Eluates were analyzed by SDS-PAGE, followed by western blotting using antiserum for the indicated proteins. Total, 1%; Eluate, 100%. (D) Immunoprecipitated FLAG-PARK2 complexes from CCCP (3 h)-treated cells were analyzed by 2D gel analysis as in (B). Asterisk indicates likely ubiquitinated forms of VDAC. (E) Immunoprecipitated FLAG-PARK2 complexes from mitochondria, after CCCP or DMSO treatment of cells, were incubated with the deubiquitinase USP2, or the buffer control, for 1 h at 37°C. Samples were split and analyzed by BN-PAGE (right panel) and SDS-PAGE (left panel), followed by western blotting with the indicated antibodies. (F) Mitochondria from FLAG-PARK2 cells treated with CCCP for 3 h were solubilized in buffer containing 2% anti-VDAC serum or the respective pre-immune serum (control). Following solubilization, protein complexes were resolved by BN-PAGE analysis (4-13%) and were subsequently immunoblotted. (G) Mitochondria from wild-type cells treated with 10 µM CCCP, or DMSO control, for 3 h were solubilized and analyzed as in (B).

Article Snippet: The deubiquitinase USP2 was purchased from BostonBiochem® (E-506).

Techniques: Labeling, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, SDS Page, Western Blot, Expressing, Two-Dimensional Gel Electrophoresis, Incubation

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Ubiquitination-Deubiquitination Balance Dictates Ligand-stimulated PTHR Sorting

doi: 10.1002/jbmr.494

Figure Lengend Snippet: (A) ROS cells were transiently transfected with HA-PTHR and treated with 100 nM PTH(1–34) or 1 μM PTH(7–34) for 1 hour. USP2 expression was assayed by immunoblot. (B) ROS cells were transfected with HA-USP2 and Flag-PTHR. After 48 hours, cells were treated with agonist as indicated. PTHR was detected using an anti-Flag primary antibody (1:1000). Values are mean ± SEM from ≥ 3 independent experiments. *p<0.05 vs. PTH(7–34).

Article Snippet: Polyclonal anti-EPS15 antibody, EPS15 shRNA and USP2 shRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Transfection, Expressing, Western Blot

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Ubiquitination-Deubiquitination Balance Dictates Ligand-stimulated PTHR Sorting

doi: 10.1002/jbmr.494

Figure Lengend Snippet: ROS cells were transfected with HA-PTHR. After 24 hours cells were transfected with shRNA-USP2 and incubated for 48 hours. Cells were then treated with 100 nM PTH(1–34) for 30 or 120 minutes. Total lysates and immunoprecipitated protein were analyzed by SDS-polyacrylamide gels and transferred to Immobilon-P membranes. Representative autoradiograms of USP2 (A) Ubiquitinated PTHR (B) and total PTHR (C) are shown. Values are mean ± SEM from ≥ 3 independent experiments. *p<0.05; **p < 0.01 vs. corresponding scrambled shRNA value.

Article Snippet: Polyclonal anti-EPS15 antibody, EPS15 shRNA and USP2 shRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Transfection, shRNA, Incubation, Immunoprecipitation

Journal: Oncotarget

Article Title: Ubiquitin-specific protease 2 decreases p53-dependent apoptosis in cutaneous T-cell lymphoma

doi: 10.18632/oncotarget.10268

Figure Lengend Snippet: Analysis of USP2 mRNA expression in paraffin-embedded biopsies from MF-plaque (n=5) and MF-tumor (n=13) a. and in quiescent lymphocytes and a panel of cell lines, Psor-2, MyLa2000, Hut-78 and SeAx c. by qPCR. Data was normalized to GAPDH and expressed as relative units (RU). Unpaired T-test was used to calculate P-value. Columns, mean (n = 3); error-bars, SD. b. USP2 protein expression in 5 MF-plaque and MF-tumor was detected by immunohistochemistry. Red color indicates USP2 expression. Bar=50μm. Arrows highlight lymphoma cells. d. Analysis of USP2 knockdown in MyLa2000 cell viability. Columns, mean (n = 8); paired T-test was used to calculate P-value, error-bars, SEM. PI-negative cells represent viable cells.

Article Snippet: For the quantification of Mdm2, USP2 and p21, 20 ng of cDNA per reaction were amplified in the presence of TaqManR universal master mix (Applied Biosystems, Foster City, CA) and TaqMan® Gene Expression Assay: Mdm2 (Hs01066930-m1), USP2 (Hs00899199-g1), p21 (Hs00355782-m1), GAPDH (Hs02758991_g1)(Applied Biosystems) (stage 1, 50°C for 2 min, stage 2, 95°C for 10 min and stage 3, 95°C for 15 s, 60°C for 1 min, repeated 45 times) in Stratagene Mx3005p (Aglicent Technologies, Santa Clara, CA).

Techniques: Expressing, Immunohistochemistry

Journal: Oncotarget

Article Title: Ubiquitin-specific protease 2 decreases p53-dependent apoptosis in cutaneous T-cell lymphoma

doi: 10.18632/oncotarget.10268

Figure Lengend Snippet: p53 wt CTCL cell line, MyLa2000, was subjected to PUVA a, b. or 5μM nutlin3a c-d. as shown in Methods. USP2 expression was measured by qPCR (a, c) and western blot (WB) (b, d) 2h-72h after the treatments. For qPCR, data was normalized to GAPDH and expressed as relative units (RU). The experiments were repeated 3 times, unpaired T-test was used to calculate P-value, error-bars, SD, *, P<0.05; **, P<0.01. For WB, Mdm2 and p53 protein level was also measured. β-actin was used as internal control, and relative protein expression levels are reported below the corresponding western blot bands. Representative data was shown.

Article Snippet: For the quantification of Mdm2, USP2 and p21, 20 ng of cDNA per reaction were amplified in the presence of TaqManR universal master mix (Applied Biosystems, Foster City, CA) and TaqMan® Gene Expression Assay: Mdm2 (Hs01066930-m1), USP2 (Hs00899199-g1), p21 (Hs00355782-m1), GAPDH (Hs02758991_g1)(Applied Biosystems) (stage 1, 50°C for 2 min, stage 2, 95°C for 10 min and stage 3, 95°C for 15 s, 60°C for 1 min, repeated 45 times) in Stratagene Mx3005p (Aglicent Technologies, Santa Clara, CA).

Techniques: Expressing, Western Blot

Journal: Oncotarget

Article Title: Ubiquitin-specific protease 2 decreases p53-dependent apoptosis in cutaneous T-cell lymphoma

doi: 10.18632/oncotarget.10268

Figure Lengend Snippet: USP2 knockdown enhances MyLa2000 apoptosis upon PUVA a. and nutlin3a b. treatment. MyLa2000 cells were transfected with USP2 siRNA or siRNA control. After 24h the cells were treated with the indicated dosage of PUVA and nutlin3a. 24h post nutlin3a and 48h post PUVA stimulation, cells were staining with annexin V and PI for flow cytometry, as described in Methods. Left: comparison of cell viability between cells with USP2 siRNA and siRNA control after treatment. Values are means of three independent experiments, paired T-test was used to calculate P-value, Error-bars, SEM. *, P<0.05; **, P<0.01. Right: The dot-plot graphs are representative of three independent experiments.

Article Snippet: For the quantification of Mdm2, USP2 and p21, 20 ng of cDNA per reaction were amplified in the presence of TaqManR universal master mix (Applied Biosystems, Foster City, CA) and TaqMan® Gene Expression Assay: Mdm2 (Hs01066930-m1), USP2 (Hs00899199-g1), p21 (Hs00355782-m1), GAPDH (Hs02758991_g1)(Applied Biosystems) (stage 1, 50°C for 2 min, stage 2, 95°C for 10 min and stage 3, 95°C for 15 s, 60°C for 1 min, repeated 45 times) in Stratagene Mx3005p (Aglicent Technologies, Santa Clara, CA).

Techniques: Transfection, Staining, Flow Cytometry

Journal: Oncotarget

Article Title: Ubiquitin-specific protease 2 decreases p53-dependent apoptosis in cutaneous T-cell lymphoma

doi: 10.18632/oncotarget.10268

Figure Lengend Snippet: a. USP2 expression is induced by nutlin3a and PUVA in Mac2a with functional p53, but not in Hut-78 and SeAx cells with impaired p53. USP2 expression was examined in Hut-78, SeAx and Mac2a cells treated with nutlin3a and PUVA as shown in figure . The experiment was repeated 3 times. Unpaired T-test was used to calculate P-value, error-bars, SD. b. Knockdown of p53 reduces induction of USP2 expression upon nutlin3a treatment. USP2 expression was checked in MyLa2000 cells with silenced p53 24h after nutlin3a treatment. Values are means of three independent experiments, paired T-test was used to calculate P-value, Error-bars, SEM. c. Hut-78 apoptosis is not increased upon USP2 knockdown after PUVA treatment. The experiment was performed and presented as described in figure . Paired T-test was used to calculate P-value, Error-bars, SEM.

Article Snippet: For the quantification of Mdm2, USP2 and p21, 20 ng of cDNA per reaction were amplified in the presence of TaqManR universal master mix (Applied Biosystems, Foster City, CA) and TaqMan® Gene Expression Assay: Mdm2 (Hs01066930-m1), USP2 (Hs00899199-g1), p21 (Hs00355782-m1), GAPDH (Hs02758991_g1)(Applied Biosystems) (stage 1, 50°C for 2 min, stage 2, 95°C for 10 min and stage 3, 95°C for 15 s, 60°C for 1 min, repeated 45 times) in Stratagene Mx3005p (Aglicent Technologies, Santa Clara, CA).

Techniques: Expressing, Functional Assay

Journal: Oncotarget

Article Title: Ubiquitin-specific protease 2 decreases p53-dependent apoptosis in cutaneous T-cell lymphoma

doi: 10.18632/oncotarget.10268

Figure Lengend Snippet: a. USP2 knockdown decreased Mdm2 expression. MyLa2000 cells were transfected with either siRNA control or USP2 siRNA for 24h, followed by treatment with 5μM nutlin3a for 24h. Western blot was used for detecting USP2, Mdm2 and p53 protein level. β-actin was used as internal control, and relative protein expression levels are reported below the corresponding western blot bands. The experiment was repeated 3 times. Representative data was shown. b. USP2 knockdown increase p53 transcription activity. The expression of p21 was used to indicate p53 transcription activity. qPCR was applied to examine p21 expression in MyLa2000 cells transfected with siUSP2/siRNA with or without nutlin3a treatment. Paired T-test was used to calculate P-value. c. A proposed model for the regulation of USP2. PUVA and nutlin3a activate p53 and promote cell apoptosis. USP2 is induced by PUVA and nutlin3a and increases cell resistance to apoptosis via modulation of Mdm2/p53 interaction, constituting an anti-apoptosis protective loop.

Article Snippet: For the quantification of Mdm2, USP2 and p21, 20 ng of cDNA per reaction were amplified in the presence of TaqManR universal master mix (Applied Biosystems, Foster City, CA) and TaqMan® Gene Expression Assay: Mdm2 (Hs01066930-m1), USP2 (Hs00899199-g1), p21 (Hs00355782-m1), GAPDH (Hs02758991_g1)(Applied Biosystems) (stage 1, 50°C for 2 min, stage 2, 95°C for 10 min and stage 3, 95°C for 15 s, 60°C for 1 min, repeated 45 times) in Stratagene Mx3005p (Aglicent Technologies, Santa Clara, CA).

Techniques: Expressing, Transfection, Western Blot, Activity Assay