Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: A The percentage of genomic alterations of USP13 in human ovarian serous adenocarcinoma (TCGA provisional databases, cBioportal). B OncoPrint showing genetic alterations of USP13 , TP53 , PI3KCA , AKT1 , AKT2 , and PTEN in human ovarian serous adenocarcinoma. C Correlation between the copy number alteration and mRNA expression of USP13 (left panel). Scatterplots of USP13 copy number versus mRNA expression in ovarian serous adenocarcinoma (right panel). D USP13 expression in different grades of ovarian cancer. E Kaplan–Meier plots of overall survival (OS), progression-free survival (PFS), and post-progression survival (PPS) for USP13 expression in patients with ovarian cancer.

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: Expressing

Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: A Schematic depicting the Trp53 flox/flox (P), Pten flox/flox (T), and Usp13 LSL/LSL (U) transgenes. Conditional overexpression of USP13 with deletion of Trp53 and Pten by Cre-mediated recombination. B Ovarian tumor incidence of Trp53 flox/flox ; Pten flox/flox (PT, n = 12) and Trp53 flox/flox ; Pten flox/flox ; Usp13 LSL/LSL (PTU, n = 18) mice after unilateral or bilateral ovarian AdCre intrabursal injection. C Kaplan–Meier curve showing overall survival of PT ( n = 6) or PTU ( n = 10) mice with bilateral AdCre intrabursal injection. P < 0.001, log-rank test. D Representative image of PT and PTU mouse after 104 days of AdCre intrabursal bilateral injection. E Ovarian tumor development in PT and PTU mice. Left side is AdCre virus injected and right side is non-injected ovary. Ov, ovary; Ut, uterus; OvT, Ovarian tumor. F Primary tumor weight (left) and tumor volume (right) were measured in each ovary of all PT and PTU mice. Unpaired Student t-test was used for statistical analyses. *** P < 0.001.

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: Over Expression, Injection

Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: Summary of ovarian tumor incidence following intrabursal injection of AdCre virus.

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: Injection

Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: Summary of metastatic tumor incidence following intrabursal injection of AdCre virus.

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: Injection

Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: A Western blot analysis of USP13, WT1, ZEB1, and Vimentin expression in ovarian primary cancer cell lines established from PT and PTU mice. B Cell proliferation of PT and PTU primary cancer cells was measured by MTS assay. Data are presented as the mean of three independent experiments. Error bars denote the standard deviation (** P < 0.01 vs PT). C Clonogenic assay of PT and PTU cells in anchorage-dependent culture for 7 days and colonies were stained by 0.05% crystal violet. Representative images are shown (right). Unpaired Student t-test was used for statistical analyses. D Wound-healing assay of PT and PTU cell lines. Experiments were performed in triplicate and representative images (left) are shown. The average wound area was calculated using Image J. Graph (right) showing relative wound healing rate to 0 h (means ± SD) of PT and PTU cells in three independent experiments. *** P < 0.001, two-tailed Student t-test . E Tumor spheroid culture of PT and PTU cell lines in Matrigel domes for 5 or 7 days, and imaged by brightfield microscopy. Black scale bars = 250 μm, white scale bars = 50 μm. F Relative mean number of spheroids ± SD ( n = 3, >6 fields/ cell line, Student t-test ) (left) or relative mean of spheroid area ± SD ( n = 3, >52 spheroid/ cell line, Student t-test ) (right). ( G ) Schematic of intraperitoneal injection of 5 × 10 5 PT or PTU cells into 7 weeks old female wild-type C57BL/6J mouse. Tumor and ascites incidence were evaluated 8 weeks post-injection and results are summarized in the table below.

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: Western Blot, Expressing, MTS Assay, Standard Deviation, Clonogenic Assay, Staining, Wound Healing Assay, Two Tailed Test, Microscopy, Injection

Journal: Oncogene

Article Title: USP13 promotes development and metastasis of high-grade serous ovarian carcinoma in a novel mouse model

doi: 10.1038/s41388-022-02224-x

Figure Lengend Snippet: Response of PT and PTU primary ovarian cancer cells to chemotherapy A Paclitaxel (48 h, 0–1 μM), B Cisplatin (48 h, 0–10 μM), or C AKT inhibitor MK-2206 (24 h, 0–10 μM). Cell viability was determined by MTT assay and calculated the relative cell viability to the vehicle (DMSO) only treated cells. Error bars, mean ± SD, n = 3. D–F PT and PTU-derived cells were treated with a combination of Spautin-1 (1 or 2 μM), a USP13 inhibitor, and MK-2206 for 24 h. G The IC50 values were summarized. Error bars, mean ± SD, with n = 3. * P < 0.05, ** P < 0.01, **** P < 0.0001 (paired Student’s t-test).

Article Snippet: For genotyping of USP13 knock-in mice, genomic DNA was isolated from tail clippings using the Cyagen Biotech (Los Angeles, CA) according to the manufacturer’s protocol.

Techniques: MTT Assay, Derivative Assay

Journal: Antioxidants

Article Title: Whey Improves In Vitro Endothelial Mitochondrial Function and Metabolic Redox Status in Diabetic State

doi: 10.3390/antiox12061311

Figure Lengend Snippet: Whey reduced the PA+HG-mediated apoptosis. ( A , B ) Representative FACS analysis of annexin V-FITC and PI-staining and immunoblotting analysis with cropped blots of ( C ) procaspase-9, ( D ) caspase-3, ( E ) PARP, ( F ) Bax, ( G ) Bcl-2, and ( H ) Bax/Bcl-2 ratio. Q1: necrotic cells; Q2: late apoptotic cells; Q3: early apoptotic cells; Q4: viable cells. Data are reported as mean ± SD of n = 3 independent experiments. M = molecular weight markers, lane 1 = Ctr, lane 2 = WH, Lane 3 = PA+HG, lane 4 = WH+PA+HG. Western blotting results are expressed as arbitrary units (AU). * p < 0.05 vs. Ctr; † p < 0.01 vs. Ctr; ‡ p < 0.001 vs. Ctr; • p < 0.05 vs. PA+HG; § p < 0.01 vs. PA+HG.

Article Snippet: Then, the membranes were immunodetected with primary antibodies against SIRT3 (1:2000, PA5-86035, Invitrogen, Waltham, MA, USA), BAX (1:500, orb216030, Biorbyt, Cambridge, UK), Bcl-2 (1:500, E-AB-15522, Elabscience Biotechnology Inc., Houston, TX, USA), COX-IV (1:2000, MA5-15078, Invitrogen, Waltham, MA, USA), caspase-3 (1:1000, 9662, Cell Signaling Technology, Danvers, MA, USA), caspase-9 (1:1000, 9508, Cell Signaling Technology, Danvers, MA, USA), PARP (1:1000, ab194586, Abcam, Cambridge, UK), cyclin D1 (1:1000, ab134175, Abcam, Cambridge, UK), cyclin E1 (1:1000, ab33911, Abcam, Cambridge, UK), SREBP1 (1:1000, Abcam, ab28481, Cambridge, UK), PPAR-α (1:1000, Elabscience Biotechnology Inc., Houston, TX, USA, E-AB-32646), PPAR-γ (1:1000, Biorbyt, orb69095, Cambridge, UK), LDHA (1:1000, ThermoFisher Scientific, PA5-27406, Waltham, MA, USA), α-tubulin (1:5000, E-AB-20036, Elabscience Biotechnology Inc., Houston, TX, USA), actin (1:3000, ab179467, Abcam, Cambridge, UK), and GAPDH (1:2000, ab9485, Abcam, Cambridge, UK).

Techniques: Staining, Western Blot, Molecular Weight