Journal: bioRxiv

Article Title: Laccaria bicolor pectin methylesterases are involved in ectomycorrhiza development with Populus tremula x Populus tremuloides

doi: 10.1101/2022.06.08.495362

Figure Lengend Snippet: Immunolocalization with pectin antibodies LM19 (recognizing de-esterified homogalacturonan, HG, (a, c)) and LM20 (recognizing methylesterified HG (b, d)) in 1µm thin cross-sections of control roots without fungus (a, b) and colonized roots (c, d) collected at 14 days after contact (DAC) (or control without fungus). L. bicolor is visualized by labelling with WGA-AF488 (green); and plant cell-wall homogalacturonan is visualized with LM19 and LM20 antibody conjugated Cy5 (magenta). White arrows indicate the epidermal layer where the difference of antibody labelling abundance is evident between ECM and control roots. Scale bar = 20 µm. Insets in images a-d show a magnified view of three adjacent epidermal cells. Fluorescence intensity ratio of LM19 and LM20 in the epidermal layer (e) and whole tissue area (f), respectively, at the indicated time-points after contact. Data was collected from ten observations per root and four to five roots per timepoint and condition. ** p-value <0.01, * p-value <0.05 with the student t-test.

Article Snippet: We indirectly quantified pectin epitopes by measuring the fluorescence probe (Cy5) intensity (mean number of pixels per unit area) conjugated with the respective primary antibody using the ‘curve spline’ function of the Zeiss ZEN blue software.

Techniques: Control, Fluorescence

Journal: Genes & Diseases

Article Title: A novel aptamer-based small RNA delivery platform and its application to cancer therapy

doi: 10.1016/j.gendis.2022.05.004

Figure Lengend Snippet: Magnesium induced assembly of nanoparticles with improved resistance to degradation and miRNA function. (A) Micelle formation under various concentrations of MgCl 2 detected by hydrophobic-incorporation of fluorescent dye (CMC-535). (B) Plots of fluorescence intensity at 535 nm under various concentrations of the delivery platform. The critical micelle concentration (CMC) was 3 × 10 -7 M. (C) Transmission electron microscopy image of the delivery platform nanoparticles using negative staining. Scale bar, 50 nm. Representative image of a single experiment. (D) The particle size distribution of the delivery platform with MgCl 2 (+Mg 2+ ) or without MgCl 2 (-Mg 2+ ) measured by dynamic light scattering. Representative image of two independent experiments. (E) Stability of each component of the miR-26a delivery platform against serum degradation in vitro . The miR-26a chimera of particle form (+Mg 2+ ) or non-particle monomer form (-Mg 2+ ) in various formats ( Fig. S3A ) was incubated with human serum for various time periods. Stability was measured by qPCR for miR-26a. (F) Functional effect of each moiety in the delivery platform for gene silencing by miR-26a. The c-Kit-targeting miR-26a chimera (1 μM) in various formats ( Fig. S3A ) with or without MgCl 2 were incubated with c-Kit + TUBO cancer cells for 2 days. The expression levels of a miR-26a target gene, Ezh2 , were measured by qPCR. Asterisks denote the significant difference compared to vehicle controls. (A, B, E, F) Data shown as mean ± SD of combined data from two independent experiments, each with duplicated samples. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Fluorescence spectroscopy was used to estimate the critical micelle concentration (CMC) of the delivery platform using a hydrophobic fluorescent probe following the manufacturer's protocol (CMC-535 detergent assay, G-Bioscience, St. Louis, MO) at 25 °C.

Techniques: Fluorescence, Concentration Assay, Transmission Assay, Electron Microscopy, Negative Staining, In Vitro, Incubation, Functional Assay, Expressing

Journal: Genes & Diseases

Article Title: A novel aptamer-based small RNA delivery platform and its application to cancer therapy

doi: 10.1016/j.gendis.2022.05.004

Figure Lengend Snippet: The pH-sensitive delivery platform induced endosomal leakage but not cytotoxicity. (A) Target specificity of the c-Kit-targeting delivery platform linked with anti-c-Kit DNA aptamer. The c-Kit-targeting aptamer-positive platform bound specifically to c-Kit + MEF cells in vitro . (B) Visualizing uptake of c-Kit-targeting miR-26a chimera in c-Kit + MEF cells over 120 min. The AF488-conjugated miR-26a chimera (Green) was incubated with the cells at 37 °C. After washing with PBS, the cells were fixed with 4% formaldehyde and images were acquired on a fluorescent microscope. Scale bar, 5 μM. (C) Destabilization of micelle particles of the miR-26a chimera under acidic pH conditions detected by hydrophobic-incorporation of fluorescent dye (CMC-535). Tween-20 is a used as a control. Asterisks denote significant differences compared to pH 7.0. (D) Free magnesium concentration in solution of the delivery platform nanoparticles incubated under various pH conditions for 1 h. Asterisks denote significant differences compared to pH 7.0. (E) Release of an endosomal trafficking fluorescent probe (10 k Dextran, red) from endosomes in c-Kit + MEF cells at 3 h after treatment with vehicle or miR-26a chimera (Top panels). Release of another fluorescent probe (Cathepsin enzymatic fluorescent substrate, Magic Red) from endosomes in c-Kit + MEF cells at 3 h after treatment with vehicle or miR-26a chimera (Bottom panels). Scale bar, 5 μM. (F) Lack of cytotoxicity of miR-26a chimera to cKit + MEF cells treated with various concentration of miR-26a chimera for 24 h as detected by LDH release assay. Asterisks denote the significant difference compared to 0 μM of miR-26a chimera treatment. (A, B, E) Representative images of two independent experiments. (C, D, F) Data shown as mean ± SD of combined data from two independent experiments, each with duplicated samples. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Fluorescence spectroscopy was used to estimate the critical micelle concentration (CMC) of the delivery platform using a hydrophobic fluorescent probe following the manufacturer's protocol (CMC-535 detergent assay, G-Bioscience, St. Louis, MO) at 25 °C.

Techniques: In Vitro, Incubation, Microscopy, Control, Concentration Assay, Lactate Dehydrogenase Assay

Journal: Cancers

Article Title: Curcumin-Dichloroacetate Hybrid Molecule as an Antitumor Oral Drug against Multidrug-Resistant Advanced Bladder Cancers

doi: 10.3390/cancers16173108

Figure Lengend Snippet: List of fine chemicals and assay kits used in this study.

Article Snippet: JC-1 dye , G-Biosciences (St. Louis, MO, USA) #786-1322.

Techniques:

Journal: Journal of Food and Drug Analysis

Article Title: Induction of G 2 /M arrest and apoptosis through mitochondria pathway by a dimer sesquiterpene lactone from Smallanthus sonchifolius in HeLa cells

doi: 10.1016/j.jfda.2016.10.005

Figure Lengend Snippet: Effect of uvedafolin on caspase-8, caspase-9, and caspase-3/-7 activities for 24 hours. The cells were labeled with fluorochrome inhibitor of caspase (FILCA) and measured by the fluorescence microplate reader (excitation: 485 nm and emission: 530 nm). Results are expressed as means ± standard deviation (SD). The data was analyzed by Duncan test using SPSS ( p < 0.05).

Article Snippet: Caspase activities were measured by using FAM-FILICA caspase inhibitor fluorescence probes (Immunochemistry Technologies, LLC, Bloomington, MN, USA) as descried in a previous report [ ].

Techniques: Labeling, Fluorescence, Standard Deviation