Journal: Molecular Plant Pathology

Article Title: The noncanonical type III secretion system of X anthomonas translucens pv. graminis is essential for forage grass infection

doi: 10.1111/mpp.12030

Figure Lengend Snippet: Genes encoding predicted type III secreted effectors found in the genome of X anthomonas translucens pv. graminis strain 29 ( X tg 29) and putative functions or homologues from other pathogens containing a type III secretion system (e.g. P seudomonas syringae , R alstonia solanacearum or Y ersinia spp.)

Article Snippet: , XTG29_00200 , + , Homology to HopR , Xanthomonas outer protein AM ( X. vesicatoria ATCC 35937) , 0.0 , 55 , 5166 , Wei et al . ( 2007 ).

Techniques: Sequencing

Journal: Molecular Plant Pathology

Article Title: The noncanonical type III secretion system of X anthomonas translucens pv. graminis is essential for forage grass infection

doi: 10.1111/mpp.12030

Figure Lengend Snippet: (a) Mean areas under the disease progress curve (AUDPCs) from four replications for L olium multiflorum genotype LmK‐01 infected with Xanthomonas translucens pv. graminis strain 29 (X tg29) deficient in genes encoding the type III secretion system (T3SS) compared with the wild‐type strain. Means with different letters are significantly different (P < 0.05) on the basis of a two‐sided t‐test using the Holm P value adjustment. The negative control treatment consisted of cutting the plants with scissors dipped in sterile physiological sodium chloride solution. (b) Colonization of L . multiflorum by X tg29 and the X tg29 strains deficient in T3SS genes. Bacterial population densities were determined for leaves of three or four different plants of LmK‐01 sampled at 6 h post‐infection (hpi) and 4, 7 and 14 days post‐infection (dpi), counting the colony‐forming units (CFU) of serial dilutions. For each time point, mean population densities and standard deviations were calculated. Means indicated with different letters are significantly (P < 0.05) different on the basis of a two‐sided t‐test using the Holm P value adjustment.

Article Snippet: , XTG29_00200 , + , Homology to HopR , Xanthomonas outer protein AM ( X. vesicatoria ATCC 35937) , 0.0 , 55 , 5166 , Wei et al . ( 2007 ).

Techniques: Infection, Negative Control

Journal: Cell Death and Differentiation

Article Title: Deubiquitinating enzyme USP33 restrains docetaxel-induced apoptosis via stabilising the phosphatase DUSP1 in prostate cancer

doi: 10.1038/s41418-019-0473-8

Figure Lengend Snippet: a C4-2B and PC3 cells were prepared for whole-cell extracts (WCE), which were immunoprecipitated (IP) with anti-USP33 or IgG as indicated. The associated DUSP1 was examined by western blot. b PC3 cells transfected with Ctrl siRNA or USP33 siRNA (USP33 RNAi #1), and USP33+/+ and USP33−/− PC3 cells were transfected with Mock, WT-USP33 or Mut-USP33 vector (si-resistance (si-res) or not) as indicated for 48 h. The indicated molecules were examined by western blot analysis. One representative experiment of three is shown. Similar results were obtained in three independent experiments. c IHC scores for DUSP1 staining in the indicated groups. Error bars represent the mean ± s.d. (n = 24, Wilcoxon signed rank test). **P < 0.01. d Representative IHC images for USP33 and DUSP1 expression in tissue microarrays. e Correlation between the IHC scores for USP33 and DUSP1 (n = 51, Pearson correlation assay).

Article Snippet: The siRNA duplexes specific for DUSP1 (sc-35937), JNK1 (sc-29380) and JNK2 (sc-39101) were from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Immunoprecipitation, Western Blot, Transfection, Plasmid Preparation, Staining, Expressing, Two-Photon Excitation Fluorescence Cross-Correlation Assay

Journal: Cell Death and Differentiation

Article Title: Deubiquitinating enzyme USP33 restrains docetaxel-induced apoptosis via stabilising the phosphatase DUSP1 in prostate cancer

doi: 10.1038/s41418-019-0473-8

Figure Lengend Snippet: a, b PC3 cells were transfected with Ctrl siRNA or USP33 siRNA (USP33 RNAi #1) (a), and USP33−/− PC3 cells were transfected with Mock or Flag-USP33 vector for 48 h (b) in the presence or absence of MG132 (10 μM) as indicated. The indicated molecules were examined by western blot analysis. c C4-2B and PC3 cells were treated with control (Ctrl) or USP33 siRNA (USP33 RNAi #1) for 48 h in the presence of MG132 (10 μM). Polyubiquitination of DUSP1 was examined by western blot (WB) analysis after immunoprecipitation (IP) with an anti-DUSP1 antibody. WCE, whole-cell extracts. d C4-2B and PC3 cells were transfected with control (Ctrl) or USP33 siRNA (USP33 RNAi #1) for 48 h in the presence of MG132 (10 μM). The K48 polyubiquitination of DUSP1 was examined by western blot (WB) assay using a K48-Ub specific antibody after immunoprecipitation (IP) with anti-DUSP1 or IgG antibody. e HEK293 cells were cotransfected with the indicated amounts (0, 0.2, 1 or 2 μg) of Flag-USP33 or Flag-USP33(mutant) and equal amounts of Myc-DUSP1 and HA-Ub (K48) vectors for 48 h in the presence of MG132 (10 μM). Then, K48-polyubiquitinated DUSP1 was examined by western blot (WB) of HA after immunoprecipitation (IP) with anti-Myc agarose. WCE, whole-cell extracts. f USP33 WT (USP33+/+) and USP33 KO (USP33−/−) PC3 cells were transfected with Mock, Flag-USP33 or Flag-USP33 (mutant) vector as indicated for 48 h in the presence of MG132 (10 μM). The K48 polyubiquitination of DUSP1 was examined by western blot (WB) with K48-Ub-specific antibody after immunoprecipitation (IP) with anti-DUSP1 or IgG antibody. MG132 in all experiments were added 6 h before cells were harvested. All cell lysates that were tested for ubiquitination in c–f were heat denatured in the presence of 1% SDS before IP. One representative experiment of three is shown. Similar results were obtained in three independent experiments.

Article Snippet: The siRNA duplexes specific for DUSP1 (sc-35937), JNK1 (sc-29380) and JNK2 (sc-39101) were from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Transfection, Plasmid Preparation, Western Blot, Immunoprecipitation, Mutagenesis

Journal: Cell Death and Differentiation

Article Title: Deubiquitinating enzyme USP33 restrains docetaxel-induced apoptosis via stabilising the phosphatase DUSP1 in prostate cancer

doi: 10.1038/s41418-019-0473-8

Figure Lengend Snippet: a PC3 cells were cotransfected with control (Ctrl) or USP33 siRNA (USP33 RNAi #1) and Mock or Myc-DUSP1 vector for 48 h and then treated with DMSO or docetaxel (10 nM) for 48 h as indicated. Annexin V+ apoptotic cells were quantified by Annexin V/PI assay. b PC3 cells were cotransfected with control (Ctrl) or USP33 siRNA (USP33 RNAi #1) and Mock or Myc-DUSP1 vector for 48 h as indicated in the presence of docetaxel (10 nM) for 24 h, and the indicated molecules were examined by western blot analysis. One representative experiment of three is shown. Similar results were obtained in three independent experiments. c USP33+/+ and USP33−/− PC3 cells were transfected with control (Ctrl) or DUSP1 siRNAs for 48 h as indicated and then treated with docetaxel (10 nM) for 48 h. Annexin V+ apoptotic cells were quantified by Annexin V/PI assay. Data and error bars in a and c represent the mean ± s.d. of triplicate samples derived from one representative experiment. Similar results were obtained in three independent experiments. *P < 0.05; ***P < 0.001; ns, not significant (one-way ANOVA followed by Bonferroni multiple comparison using PRISM software). d USP33+/+ and USP33−/− PC3 cells were transfected with control (Ctrl) or DUSP1 siRNA for 48 h as indicated and then treated with docetaxel (10 nM) for 24 h, and the indicated molecules were examined by western blot analysis. One representative experiment of three is shown. Similar results were obtained in three independent experiments.

Article Snippet: The siRNA duplexes specific for DUSP1 (sc-35937), JNK1 (sc-29380) and JNK2 (sc-39101) were from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Plasmid Preparation, Western Blot, Transfection, Derivative Assay, Software

Journal: Cell Death and Differentiation

Article Title: Deubiquitinating enzyme USP33 restrains docetaxel-induced apoptosis via stabilising the phosphatase DUSP1 in prostate cancer

doi: 10.1038/s41418-019-0473-8

Figure Lengend Snippet: a, b Tumour volume was measured 3 days after the indicated treatments. When the PC3 tumour volume reached 70–100 mm3 (day 7), the tumours with the required size (n = 4) were treated as indicated. Black arrows indicate the siRNA injections. Docetaxel was intraperitoneally injected at 10 mg/kg once every 5 days from the time the tumour reached the required size (70–100 mm3) for a total of three times. c Representative IHC images of USP33 and DUSP1 protein staining in tumour tissues. d, e Tumour volume was measured 5 days after the indicated treatments. When USP33+/+ and USP33−/− PC3 tumour volumes reached 70–100 mm3 (day 10), the tumours with required sizes (n = 3) were treated as indicated. Docetaxel was intraperitoneally injected at 10 mg/kg once every 5 days for a total of three times. Black arrows indicate the docetaxel injections. Error bars in b and e represent the mean ± s.d. ***P < 0.001 (one-way ANOVA followed by Bonferroni multiple comparison using PRISM software). f Protein was extracted from each tumour tissue, and the indicated molecules were examined by western blot analysis. #1–#3 is the number of the tumours in d.

Article Snippet: The siRNA duplexes specific for DUSP1 (sc-35937), JNK1 (sc-29380) and JNK2 (sc-39101) were from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Injection, Staining, Software, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Dual-specificity phosphatase 1 interacts with prohibitin 2 to improve mitochondrial quality control against type-3 cardiorenal syndrome

doi: 10.7150/ijms.90484

Figure Lengend Snippet: DUSP1 overexpression sustains heart function after CRS-3. DUSP1 Tg and WT mice were subjected to 30 minutes of bilateral renal artery ischemia, followed by 72 hours of reperfusion to induce CRS-3. A-G. Echocardiography was used to evaluate heart function in DUSP1 Tg and WT mice upon CRS-3. H-M. Single cardiomyocytes were isolated from DUSP1 Tg and WT mice, and their contractile/relaxation capacities were recorded. N. H&E staining was used to observe the cardiac structure following CRS-3. O-R. Serum samples were collected from DUSP1 Tg and WT mice, and the concentrations of TnT, CK-MB, BNP and LDH were determined using ELISAs. *p<0.05.

Article Snippet: DUSP1 siRNAs (#sc-35937; Santa Cruz Biotechnology, Inc.) were transfected into the HL-1 cells with Lipofectamine™ 3000 reagent, as described in our previous study .

Techniques: Over Expression, Isolation, Staining

Journal: International Journal of Medical Sciences

Article Title: Dual-specificity phosphatase 1 interacts with prohibitin 2 to improve mitochondrial quality control against type-3 cardiorenal syndrome

doi: 10.7150/ijms.90484

Figure Lengend Snippet: DUSP1 reduces the inflammatory response, oxidative stress and mitochondrial injury in the heart during CRS-3. DUSP1 Tg and WT mice were subjected to 30 minutes of bilateral renal artery ischemia, followed by 72 hours of reperfusion to induce CRS-3. A-C. RNA was collected from DUSP1 Tg and WT hearts, and qPCR was used to evaluate the transcription of IL-6 , TNFα and MCP1 . D, E. Double immunofluorescence staining was used to measure Gr-1-positive neutrophil accumulation in the myocardium. TnT was used to visualize myocardial fibers, and 4′,6-diamidino-2-phenylindole (DAPI) was used to stain the nuclei. F-H. ELISAs were used to evaluate the concentrations of glutathione (GSH), SOD and CAT in the heart. I, J. Cardiomyocytes were isolated from DUSP1 Tg and WT mice, and ROS levels were determined using DCFH-DA. K. ATP production in the myocardium was determined with an ELISA. L-M. An immunofluorescence assay was used to determine the mitochondrial membrane potential in cardiomyocytes isolated from DUSP1 Tg and WT mice after CRS-3. *p<0.05.

Article Snippet: DUSP1 siRNAs (#sc-35937; Santa Cruz Biotechnology, Inc.) were transfected into the HL-1 cells with Lipofectamine™ 3000 reagent, as described in our previous study .

Techniques: Double Immunofluorescence Staining, Staining, Isolation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Membrane

Journal: International Journal of Medical Sciences

Article Title: Dual-specificity phosphatase 1 interacts with prohibitin 2 to improve mitochondrial quality control against type-3 cardiorenal syndrome

doi: 10.7150/ijms.90484

Figure Lengend Snippet: DUSP1 overexpression maintains cardiac mitochondrial quality control during CRS-3. DUSP1 Tg and WT mice were subjected to 30 minutes of bilateral renal artery ischemia, followed by 72 hours of reperfusion to induce CRS-3. A-D. RNA was collected from DUSP1 Tg and WT hearts, and qPCR was used to evaluate the transcription of Drp1 , Fis1 , Mfn2 and Opa1 . E-G. Immunofluorescence was used to stain the mitochondria of cardiomyocytes isolated from DUSP1 Tg and WT mice. The average length of the mitochondria and the proportion of fragmented mitochondria were recorded. H-J. RNA was collected from DUSP1 Tg and WT hearts, and qPCR was used to evaluate the transcription of Parkin , Fundc1 and Beclin1 . K-M. RNA was collected from DUSP1 Tg and WT hearts, and qPCR was used to evaluate the transcription of PGC1α , Nrf2 and Tfam . *p<0.05.

Article Snippet: DUSP1 siRNAs (#sc-35937; Santa Cruz Biotechnology, Inc.) were transfected into the HL-1 cells with Lipofectamine™ 3000 reagent, as described in our previous study .

Techniques: Over Expression, Immunofluorescence, Staining, Isolation

Journal: International Journal of Medical Sciences

Article Title: Dual-specificity phosphatase 1 interacts with prohibitin 2 to improve mitochondrial quality control against type-3 cardiorenal syndrome

doi: 10.7150/ijms.90484

Figure Lengend Snippet: DUSP1 binds to PHB2 to sustain its phosphorylation in the heart during CRS-3. DUSP1 Tg and WT mice were subjected to 30 minutes of bilateral renal artery ischemia, followed by 72 hours of reperfusion to induce CRS-3. A. RNA was collected from DUSP1 Tg and WT hearts, and qPCR was used to evaluate the transcription of PHB2 . B, C. Proteins were isolated from DUSP1 Tg and WT hearts, and Western blotting was used to analyze PHB2 expression. D, E. Molecular docking analysis was used to evaluate the potential binding sites between DUSP1 and PHB2. F, G. Co-immunoprecipitation was used to analyze the crosslinks between DUSP1 and PHB2. H, I. HL-1 cells were transfected with siRNA against DUSP1, and the phosphorylation of PHB2 was determined via Western blotting. *p<0.05.

Article Snippet: DUSP1 siRNAs (#sc-35937; Santa Cruz Biotechnology, Inc.) were transfected into the HL-1 cells with Lipofectamine™ 3000 reagent, as described in our previous study .

Techniques: Isolation, Western Blot, Expressing, Binding Assay, Immunoprecipitation, Transfection