Journal: Reproduction (Cambridge, England)

Article Title: Development of polarity-reversed endometrial epithelial organoids

doi: 10.1530/REP-23-0478

Figure Lengend Snippet: Generation and characterization of apical-out organoids. (A) Schematic of approach to generate apical-out endometrial epithelial organoids (AO-EEO or AO); (B) Characterization of AO. (i–ii) cartoons of epithelial cells in apical-in organoids (AI) exhibiting apical surface facing the lumen or in AO exhibiting apical surface exposed to the outer environment; (iii–iv) bright field images of organoids grown in Cultrex BME or in suspension culture for eight days showing gross morphology; scale bars, 100 µm; (v–vi) Immunofluorescence staining of apical surface marker zonula occludens 1 (ZO1) on paraffin sections of AI or AO; scale bars, 20 µm and 60 µm for v and vi respectively. Sections were counterstained with Hoechst to detect DNA; Areas outlined in red boxes in v and vi magnified to demonstrate apical and basal surfaces, separated by vertical line; (vii–viii) scanning electron microscopy (SEM) images of AI or AO showing surface microarchitecture; scale bars, 50 µm. (C) Time-dependent screening of organoids flipping from day 0 to day 8 detected by ZO1 immunofluorescence (i–iv and vi–ix), categorized as apical-in, apical-out, or mixed polarity and quantified in (v, x); scale bars, 50 µm (i, vi) and 150 µm (ii–iv and vii–ix). Schematic in A created with Biorender.com.

Article Snippet: Those cells were washed and subsequently resuspended in Cultrex BME (Cat. No. 343300501, R&D Systems) and plated in 20–25 μL droplets of Cultrex.

Techniques: Suspension, Immunofluorescence, Staining, Marker, Electron Microscopy

Journal: Journal of Functional Foods

Article Title: Procyanidin B2 ameliorates endothelial dysfunction induced by nicotine via the induction of tetrahydrobiopterin synthesis

doi: 10.1016/j.jff.2022.105306

Figure Lengend Snippet: Fig. 8. Inhibition of PPARδ abrogates the effects of PCB2 on vasorelaxation. (A) C57BL/6J mouse thoracic artery rings were treated with 1 μM GSK0660 for 1 h followed by incubation with 10 μM PCB2 for 12 h before exposure to 1 μM nicotine for 24 h. ACh-induced relaxations were determined (n = 5). Data represent the mean ± SEM (**, p < 0.01 versus vehicle; #, p < 0.05 versus nicotine; †, p < 0.05 versus nicotine + PCB2; two-way ANOVA with post Bonferroni test). HUVECs were treated with 1 μM GSK0660 for 1 h followed by incubation with 10 μM PCB2 for 12 h before exposure to 1 μM nicotine for 24 h. (B) The mRNA levels of DHFR and GCH1 were examined using qRT-PCR. (C) The protein levels of DHFR, GCH1, p-eNOS and eNOS were detected by western blotting (n = 3). BAECs were treated with 1 μM GSK0660 for 1 h followed by incubation with 10 μM PCB2 for 12 h before exposure to 1 μM nicotine for 24 h. (D) BH4 concentration was detected using ELISA (n = 5). (E) ROS generation was detected with DHE, and the mean fluorescence intensity was evaluated (n = 3). Scale bar: 50 μm. (F) NO levels were detected with DAF-FM DA, and the mean fluorescence intensity was evaluated (n = 3). Scale bar: 100 μm. Data represent the mean ± SD (*, p < 0.05; **, p < 0.01; one-way ANOVA with post Bonferroni test).

Article Snippet: GSK0660 (Cat. No. 3433) and 1400 W (Cat. No. 1415) were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Inhibition, Incubation, Quantitative RT-PCR, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Fluorescence