Journal: ACS Nano

Article Title: Immune Modulatory Oxysterols Produced from Cholesterol-Containing Lipid Nanoparticles Regulate Tumor Growth

doi: 10.1021/acsnano.5c22020

Figure Lengend Snippet: LNP-oxysterols reduce tumor cell viability by inducing late apoptosis and necrosis in vitro. A) TC-1 tumor cell viability at 24, 48, and 72 h by MTT assay. B–E) Apoptosis and necrosis assessed at 24 h by propidium iodide (PI)/annexin V staining. Each data point represents a biological replicate. Statistical tests were performed by one-way ANOVA compared to vehicle without correction for multiple comparisons, where * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. F) Representative flow cytometry plots, where PI – /annexin V – are viable cells, PI + /annexin V + are cells in late apoptosis, PI – /annexin V + are cells in early apoptosis, and PI + /annexin V – are cells in necrosis. G) Pie charts of the percentage of viable, early apoptotic, late apoptotic, and necrotic cells.

Article Snippet: For proliferation studies, TC-1 tumor cells or BMDMs were plated in triplicate in 96-well plates, incubated at 37 °C with 5% CO 2 overnight for acclimation, and then treated with 55.7 μM LNP-cholesterol or LNP-oxysterols for 24, 48, and 72 h. Cytotoxicity and cell proliferation were assessed using the MTT assay kit (Cat. #4890050K, R&D Systems), and absorbance at 570 nm was measured using a Cytation 5 plate reader.

Techniques: In Vitro, MTT Assay, Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Long Non-Coding RNA 74687 Regulates Meiotic Progression and Gonadal Development in Rainbow Trout ( Oncorhynchus mykiss ) via the miR-15a-5p– ccne1 Regulatory Axis

doi: 10.3390/ijms26168036

Figure Lengend Snippet: 5-ethynyl-2′-deoxyuridine (EdU) assay of effect of ccne1 on proliferative capacity of rainbow trout gonadal (RTG)-2 cells. ( A ) EdU results for RTG-2 cells under different transfection conditions. ( B ) Quantification of lnc74687.3 ’s effects on proliferation by ImageJ (ImageJ, 1.53q). ( C ) Quantification of ccne1 ’s effects on proliferation by ImageJ. Data are expressed as mean ± SD ( n = 3), with differences indicated by asterisks (** p < 0.01, * p < 0.05).

Article Snippet: Cell proliferation was assessed using an EdU Cell Proliferation Kit with Alexa Fluor 555 (Beyotime, Shanghai, China) following the manufacturer’s instructions with minor optimisation.

Techniques: EdU Assay, Transfection

Journal: Cell Death and Differentiation

Article Title: Deubiquitinase USP39 and E3 ligase TRIM26 balance the level of ZEB1 ubiquitination and thereby determine the progression of hepatocellular carcinoma

doi: 10.1038/s41418-021-00754-7

Figure Lengend Snippet: A The expression of USP39 mRNA was analyzed by RT-PCR in SK-hep-1 and HepG2 HCC cells infected with shRNAs. B Western blotting analysis of USP39 protein level in SK-hep-1 and HepG2 cells infected with shRNAs. C – E The effect of USP39 on HCC cells (SK-hep-1) proliferation was determined by MTT assays ( C ) at different time points and colony formation ( D ). The colony counts were normalized to the control and expressed as a percentage, and results are represented in the bar graph ( E ). F – I Representative images of HCC cell migration ability as shown by wound-healing assays ( F – G ) and migration assay ( H – I ). Student’s t test: * P < 0.05; ** P < 0.01; *** P < 0.001. All the data are representative of at least three independent experiments and presented as the means ± SD.

Article Snippet: HCC cell viability was detected using a cell proliferation assay kit (Beyotime, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, Western Blot, Control, Migration

Journal: Cell Death and Differentiation

Article Title: Deubiquitinase USP39 and E3 ligase TRIM26 balance the level of ZEB1 ubiquitination and thereby determine the progression of hepatocellular carcinoma

doi: 10.1038/s41418-021-00754-7

Figure Lengend Snippet: A – D RT-PCR and western blotting indicated that the mRNA expression levels of USP39 and TRIM26 ( A , C ) and the protein level of ZEB1 ( B , D ) in SK-hep-1 cells co-translated with USP39 and TRIM26 expressing plasmids. ( E , G ) The mRNA levels of USP39 and TRIM26 were analyzed by RT-PCR in SK-hep-1 HCC cells. F , H Effect of USP39-knockdown and TRIM26 silencing on the protein level of ZEB1 in SK-hep-1 HCC cell determined by western blotting. I Effect of overexpression of USP39 and TRIM26 on the ZEB1 ubiquitination in SK-hep-1 cells. J Effect of USP39-knockdown and TRIM26 silencing on the cell proliferation of SK-hep-1 assessed by MTT assay. Student’s t test: * P < 0.05; ** P < 0.01; *** P < 0.001. All the data are representative of at least three independent experiments and presented as the means ± SD.

Article Snippet: HCC cell viability was detected using a cell proliferation assay kit (Beyotime, China).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Knockdown, Over Expression, Ubiquitin Proteomics, MTT Assay

Journal: Cell Death and Differentiation

Article Title: Deubiquitinase USP39 and E3 ligase TRIM26 balance the level of ZEB1 ubiquitination and thereby determine the progression of hepatocellular carcinoma

doi: 10.1038/s41418-021-00754-7

Figure Lengend Snippet: A The effect of USP39 and TRIM26 in HCC cell proliferation in vivo was determined by xenograft assays. USP39 and TRIM26 knockdown SK-hep-1 cells were respectively injected into flanks of BALB/c nude mice. After 30 days, tumors were isolated and photographed. B Tumor volumes were calculated. C Tumor weight. D , E Levels of USP39, TRIM26, and ZEB1 were analyzed by RT-PCR ( D ) and western blotting ( E ). F The expression levels of USP39, TRIM26, ZEB1 and Ki67 in tumors of different groups by IHC (original magnification, ×40; inlet, ×10). G , H Representative images showed the tumors metastasis of different groups by whole-body bioluminescence imaging ( G ) and lung metastases ( H ). The number of nodules in the lung was counted and statistically analyzed. Student’s t test: * P < 0.05; ** P < 0.01; *** P < 0.001. All the data are representative of at least three independent experiments and presented as the means ± SD.

Article Snippet: HCC cell viability was detected using a cell proliferation assay kit (Beyotime, China).

Techniques: In Vivo, Knockdown, Injection, Isolation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Imaging