Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Tenulin and isotenulin inhibit P-glycoprotein function and overcome multidrug resistance in cancer cells

doi: 10.1016/j.phymed.2018.09.008

Figure Lengend Snippet: Reversal ability of tenulin and isotenulin on MDR cancer cell line. (A) ABCB1 mRNA expression levels were quantified by real-time RT-PCR in HeLaS3 and MDR KB-vin cancer cell line with or without tenulin or isotenulin 72h treatment. (B) Apoptotic cells detection after tenulin or isotenulin 72 h treatment in HeLaS3 and KB-vin cancer cells. Apoptotic and necrotic cells were determined by Annexin V (FITC) plus propidium iodide (PI) double staining and flow cytometry. Quadrant diagrams represent cell distribution in early apoptosis (Q1), apoptosis (Q2), live (Q3), and dead (Q4). Data presented as mean ± SE of at least three experiments, * denotes p < 0.05 compared with control group.

Article Snippet: Parental and multi-drug resistant cancer cell lines Human cervical carcinoma cell line HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Expressing, Quantitative RT-PCR, Double Staining, Flow Cytometry, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Tenulin and isotenulin inhibit P-glycoprotein function and overcome multidrug resistance in cancer cells

doi: 10.1016/j.phymed.2018.09.008

Figure Lengend Snippet: Effects of tenulin and isotenulin on cytotoxicity of chemotherapeutic drugs in HeLaS3 and KB-vin cells.

Article Snippet: Parental and multi-drug resistant cancer cell lines Human cervical carcinoma cell line HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques:

Journal: bioRxiv

Article Title: Enviromic-based Kernels Optimize Resource Allocation with Multi-trait Multi-environment Genomic Prediction for Tropical Maize

doi: 10.1101/2021.06.11.448049

Figure Lengend Snippet: Gain per cost × 10 −3 (per 10,000 dollars invested) for the HEL and USP datasets, comparing the two optimization kernels (GET and GWT) and the standard scenario (MTMET CV2, TRN = 70%). The cost includes the phenotyping of TRN (3 USD per trait per plot) and the whole dataset’s genotyping (20 USD per sample)

Article Snippet: Parental inbred lines from HEL and USP datasets were genotyped with an Affymetrix® Axiom® Maize Genotyping SNP array of 616 K ( ).

Techniques:

Journal: The Journal of Cell Biology

Article Title: WDR62 regulates spindle dynamics as an adaptor protein between TPX2/Aurora A and katanin

doi: 10.1083/jcb.202007167

Figure Lengend Snippet: Characterization of WDR62 knock-in and knockout cell lines. (A–C) Characterization of WD62 knock-in HeLa cell line. (A) Schematic illustration of primer sets and the expected PCR products. PCR genotyping (B) and Western blotting analysis (C) showing that the WDR62-GFP-Strep knock-in HeLa cell line used in this study is heterozygous. BSD, blasticidin S deaminase. (D) Western blotting with the indicated antibodies in control, katanin p80 knockout (KO), and WDR62 knockout HeLa cells. (E) Sequencing results of WDR62 knockout HeLa cell line used in this study. 1 nucleotide (nt) insertion will result in p.V65VfsX13. sgRNA, single-guide RNA. (F) Frequency of multipolarity in control or the indicated knockout HeLa cells in prometaphase or metaphase. Control, n = 224 cells; p80 knockout, n = 189; WDR62 knockout, n = 211. (G–I) Quantification of mitotic index (G), NuMA total intensity (H), and γ-tubulin total intensity (I) in control, p80 knockout, and WDR62 knockout HeLa cells. For mitotic index, n = 3 experiments, control, 1,363 cells; p80 knockout, 1,708 cells; WDR62 knockout, 1,385 cells. For NuMA intensity, n = 70 spindle poles in all conditions. For γ-tubulin intensity, n = 74 centrosomes in all conditions. (J) Western blotting with the indicated antibodies in control, katanin p80 knockout, and WDR62 knockout U2OS stable cell lines expressing PA-GFP–α-tubulin. Both knockouts are from a mixed population of cells after transient transfection with PX459 bearing single guide RNA) followed by drug selection, rather than from single cell cloning. (K and M) Immunofluorescence staining for KIF2A, γ-tubulin, and DAPI (K) and quantification of KIF2A intensity at spindle poles (M) in control and WDR62 knockout HeLa cells. n = 50 spindles poles for both conditions. (L and N) Immunofluorescence staining for γ-tubulin and DAPI (L) and quantification of KIF2C intensity at centrosomes (N) in control and WDR62 knockout KIF2C-GFP-Strep knock-in HeLa cells. The WDR62 knockout is from a mixed population of cells after transient transfection with PX459 bearing single guide RNA followed by drug selection, rather than from single cell cloning. n = 50 spindles poles for both conditions. (O and P) Western blotting (O) and PCR genotyping (P) showing that the KIF2C-GFP-Strep knock-in HeLa cell line used in L is homozygous. Red asterisk denotes a nonspecific band detected by the KIF2C antibody. (Q) Western blotting with the indicated antibodies in control and WDR62 knockout KIF2C-GFP-Strep knock-in HeLa cells. The WDR62 knockout is from a mixed population of cells after transient transfection with PX459 bearing single guide RNA followed by drug selection, rather than from single cell cloning. Scale bars, 2 µm. Data represent mean ± SD.

Article Snippet: We first generated the Tet-3xHA-OsTIR1-puro parental HeLa cell line. pMK243 plasmid (Addgene; #72835) was modified to generate the donor construct for inducible expression of 3xHA-OsTIR1 at AAVS1 locus.

Techniques: Knock-In, Knock-Out, Western Blot, Control, Sequencing, Stable Transfection, Expressing, Transfection, Selection, Cloning, Immunofluorescence, Staining