Journal: Molecular plant

Article Title: The WAK-like protein RFO1 acts as a sensor of the pectin methylation status in Arabidopsis cell walls to modulate root growth and defense.

doi: 10.1016/j.molp.2023.03.015

Figure Lengend Snippet: Figure 6. RFO1-GFP particle dwell time at the PM is altered by the perturbation of cell wall pectin methylation. (A) Representative spinning-disc confocal images (top) of a 5-day-old root elongating epidermal cell and kymographs (bottom) along the length of the dashed yellow lines from the corresponding video (n = 50 frames, 1 frame/s) of RFO1-GFP (rfo1-1 pRFO1RFO1-GFP) and RFO1-GFP iPMEIox (rfo1-1 pRFO1RFO1-GFP iPMEIox) seedlings treated with 12 mM estradiol for 2 h. RFO1-GFP iPMEIox seedlings were co-treated with Mock () or 10 mg/ml of dmPectin (+). Unless specified, all data derived from 180-s videos taken at 1 frame/s. Scale bar, 5 mm. (B) Dwell time frequencies of RFO1-GFP particles at the PM as depicted in (A). Means (m) are shown by dashed lines. For each condition, data represent N > 12 000 particles from a minimum of three cells per root and three roots. (C) Average RFO1-GFP particle dwell times at the PM. (D) Quantification of the total number of RFO1-GFP particles at the PM (left), and particles with dwell times >50 s (middle) and >100 s (right). (E) Average instantaneous diffusion coefficients of RFO1-GFP particles at the PM. (C–E) Data represent the mean ± SE of N > 25 cells per condition from three independent replicates using three cells per root, three roots per condition, and tracking R1072 particles/cell, obtained from videos as described in (A). One-way ANOVA with Dunnett’s multiple-comparison test, **p < 0.01, ****p < 0.0001.

Article Snippet: RFO1-GFP particles were imaged with a CSU-W1 Yokogawa spinning-disk head fitted to a Nikon Eclipse Ti-E-inverted microscope with a CFI PlanApo 3 100 NA 1.40 oil immersion objective and two iXon Ultra EM-CCD cameras (Andor, GB); a 31.2 lens between the spinning disk and camera was used.

Techniques: Methylation, Derivative Assay, Diffusion-based Assay, Comparison

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 2. Tumor conditioning induces macrophage CD163 and IL-10 gene expression. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their gene expression was compared to control macrophages by RT-qPCR. A-F: Presents gene expression of polarization markers (CD163, IL-10, and TNF-α) and phagocytosis checkpoints (SIRPα, LILRB1, and Siglec-10) in tumor-associated macrophages compared to controls. Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 4.

Article Snippet: The plates were read at 450/ 620 nm and a standard curve ranging from 0.625 to 8 μg/L was prepared using recombinant human LILRB1 (R&D systems, 8989-T2). sCD206 (samples diluted 1:5) [34], and sSIRPα (samples diluted 1:10) [35] were measured as previously reported in detail.

Techniques: Gene Expression, Derivative Assay, Generated, Control, Quantitative RT-PCR, Transformation Assay

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 3. Tumor cell conditioning induces CD163, CD206, CD80 and LILRB1 on the membrane protein level. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their membrane protein expression compared to control macrophages by flow cytometry. The gating strategy is presented in Supplementary fig. 1. A-G: Presents median fluorescence intensities (MFI) of polarization markers (CD163, CD206, and CD80) and MFI values and percent macrophages positive for phagocytosis checkpoints (LILRB1, and Siglec-10). Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 9.

Article Snippet: The plates were read at 450/ 620 nm and a standard curve ranging from 0.625 to 8 μg/L was prepared using recombinant human LILRB1 (R&D systems, 8989-T2). sCD206 (samples diluted 1:5) [34], and sSIRPα (samples diluted 1:10) [35] were measured as previously reported in detail.

Techniques: Membrane, Derivative Assay, Generated, Expressing, Control, Flow Cytometry, Fluorescence, Transformation Assay