Journal: Cell chemical biology

Article Title: TREM2 is thyroid hormone regulated making the TREM2 pathway druggable with ligands for thyroid hormone receptor.

doi: 10.1016/j.chembiol.2021.07.014

Figure Lengend Snippet: Figure 4. T3 and sobetirome stimulate and NH-3 blocks phagocytosis by microglia (A–D’) C57BL/6 mouse primary microglia cells in culture were treated with DMSO vehicle (A + A’), 10 nM T3 (B + B’), 1 mM sobetirome (C + C’), or 2 mM NH-3 (D + D’) for 24 h before 2 mm diameter fluorescent beads (100/cell) were introduced 2 h before cells were fixed and stained for CD11B (green). (E) Elongated morphology of an unactivated/ramified microglia cell. (F) Morphology of an activated microglia cell upon treatment with sobetirome (see the supplemental information). (G) Three-dimensional view down three separate axes for visualization of beads inside the uppermost cell in the sobetirome treatment group (image generated using Imaris Section function). (H) Quantification of phagocytosed beads per treatment group. Scale bars, 20 mm (A–D), 10–20 mm (A’–D’), and 8–10 mm (E–G), as noted. Statistical significance was determined by a two-tailed, unpaired t test for comparisons between vehicle and group then were plotted together. Asterisks represent significant difference from vehicle unless otherwise noted: *p % 0.05, **p % 0.01, ***p % 0.001. All graphs show mean ± SEM.

Article Snippet: The cells were then permeabilized with PBS containing 0.05% saponin (saponin was included in all subsequent incubations and washes) and stained for microglial marker CD11b using monoclonal rat anti-mouse CD11b (1:200 dilution, AbD Serotec, #MCA711) in conjunction with Alexa Fluor 488-conjugated donkey anti-mouse secondary antibody (1:400 dilution,Thermo Fisher, A21202).

Techniques: Staining, Generated, Two Tailed Test

Journal: Molecular cancer

Article Title: The circular RNA circDLG1 promotes gastric cancer progression and anti-PD-1 resistance through the regulation of CXCL12 by sponging miR-141-3p.

doi: 10.1186/s12943-021-01475-8

Figure Lengend Snippet: Fig. 4 CircDLG1 promotes tumor progression by increasing the infiltration of MDSCs. a C57BL/6 mice were injected with MFC-sh-NC or MFC-sh-circDLG1 cells, and the tumor volume and tumor metastasis are shown. The mice were sacrificed at 4 weeks after injection. Left: the tumor size was measured, and the tumor volume was calculated every 4 days (n = 10 mice per group), **P < 0.05, two-way ANOVA. Right: the lungs of the mice were removed and paraffin embedded, consecutive sections (4 μm) were made and stained with hematoxylin-eosin, and the micrometastases in the lungs were examined and counted under a dissecting microscope (n = 10 mice per group), **P < 0.05, Student’s t-test. All experiments were repeated three times. Scale bar, left: 5 mm; right: 100 μm. b BALB/c nude mice were injected with MFC-sh-NC or MFC-sh-circDLG1 cells. The mice were sacrificed at 4 weeks after injection. Left: the tumor size was measured, and the tumor volume was calculated every 4 days (n = 10 mice per group). ns, no significance, two-way ANOVA. Right: The lungs of the mice were removed and paraffin embedded. Consecutive sections (4 μm) were made and stained with hematoxylin-eosin, and the micrometastases in the lungs were examined and counted under a dissecting microscope (n = 10 mice per group). ns, no significance, Student’s t-test. All experiments were repeated three times. Scale bar, left: 5 mm; right: 100 μm. c IHC analysis was performed using subcutaneous tumors for CD4, CD8, Gr-1, and CD11b (Gr-1 and CD11b are mouse MDSC markers). HPF, high-power visual field. ns, no significance; **P < 0.05. Student’s t-test. d Flow cytometric analysis of immune cells in subcutaneous tumors. **P < 0.05, Student’s t-test; the experiment was repeated three times. e The expression of CD33 (a marker of human MDSCs) was detected by IHC in human gastric cancer tissues. Scale bar, 50 μm. f The correlation between circDLG1 expression measured by qRT–PCR and CD33+ cells. n = 30, *P < 0.05, Pearson correlation analysis

Article Snippet: The following antibodies were used: anti-mouse IFNγ (BD Biosciences), anti-mouse CD45 (BD Biosciences), anti-mouse Gr-1 (TONBO Biosciences), anti-mouse Ly6G (TONBO Biosciences), anti-mouse ly6C (BD Biosciences), anti-mouse CD11b (TONBO Biosciences), anti-mouse F4/80 (BioLegend), anti-human CD11b (TONBO Biosciences), anti-human CD33 (BD Biosciences), and anti-human HLA-DR (BD Biosciences).

Techniques: Injection, Staining, Microscopy, Expressing, Marker, Quantitative RT-PCR