Journal: bioRxiv

Article Title: RORγ bridges cancer-driven lipid dysmetabolism and myeloid immunosuppression

doi: 10.1101/2023.11.19.567414

Figure Lengend Snippet: MN/MCA1 mice were fed NCD or HCD. a-d, Frequency of RORγ + CCR2 + cells and relative RORγ expression levels in: a, M-MDSCs from tumors (n=6); b, lung IMs (n=4), AMs (n=4), and M- MDSCs (n=5); c, BM M-MDSCs (n=5); d, Spleen M-MDSCs (n=5). e, Frequency of RORγ + CCR2 - cells and relative RORγ expression levels in lung AMs (n=4), BM or spleen M-MDSCs (n=5). f, Frequency of RORγ + CMPs and GMPs cells and relative RORγ expression levels (n=4). g, HEK293 cells were co-transfected with CMV-RORγ and IL17A_prom-Luc plasmids. Plasmid 1: RORγ gene is constitutively expressed under the control of CMV promoter. Plasmid 2: Luciferase reporter gene is under the control of the minimal promoter of Il17a gene, whose transcription is driven by RORγ. h-j, HEK293 cells, co-transfected with plasmids 1 and 2, were stimulated with SR0987, desmosterol, cholesterol, or LDL ( h ), or with sera from NCD- or HCD-fed MN/MCA1 TB or TF mice (n=4) ( i ), or exposed to single or combinations of cholesterol, SR2211 and TCM ( j ). Luciferase activity was quantified as relative light units (RLU). Data are representative of five ( a, b, d ), two ( c, e, h-j ), or three ( f ) independent experiments. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 between selected relevant comparisons. a-f, Unpaired two-tailed t -test; h-j, One-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: A plasmid vector constitutively expressing the Rorc gene under the control of the CMV promoter (pCMV6- Rorc ) (Origene, No. MR222309) was co-transfected with a plasmid expressing the luciferase gene ( Luc ) under the control of the minimal promoter of the Il17a gene (minIL17prom-Luc) (Addgene, No. 20124).

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Luciferase, Activity Assay, Two Tailed Test

Journal: International journal of systematic and evolutionary microbiology

Article Title: Arthrobacter bussei sp. nov., a pink-coloured organism isolated from cheese made of cow's milk.

doi: 10.1099/ijsem.0.004125

Figure Lengend Snippet: Fig. 1. Pigmentation of strain KR32T (a) and Arthrobacter agilis DSM 20550T (c) at 10 °C growth temperature after 7 days and of strain KR32T (b) and Arthrobacter agilis DSM 20550T (d) at 30 °C growth temperature after 72 h in TSB.

Article Snippet: The reference strains A. agilis DSM 20550T and Arthrobacter globiformis DSM 20124T were obtained from Leibniz- Institut– Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) for comparative studies.

Techniques:

Journal: International journal of systematic and evolutionary microbiology

Article Title: Arthrobacter bussei sp. nov., a pink-coloured organism isolated from cheese made of cow's milk.

doi: 10.1099/ijsem.0.004125

Figure Lengend Snippet: Fig. 1. Pigmentation of strain KR32T (a) and Arthrobacter agilis DSM 20550T (c) at 10 °C growth temperature after 7 days and of strain KR32T (b) and Arthrobacter agilis DSM 20550T (d) at 30 °C growth temperature after 72 h in TSB.

Article Snippet: The reference strains A. agilis DSM 20550T and Arthrobacter globiformis DSM 20124T were obtained from Leibniz- Institut– Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) for comparative studies.

Techniques:

Journal: Science signaling

Article Title: Spatially-structured cell populations process multiple sensory signals in parallel in intact vascular endothelium

doi: 10.1126/scisignal.aar4411

Figure Lengend Snippet: (A) Representative experiment showing the effect of the P2Y1 antagonist MRS2179 (10 μM) and P2Y2 antagonist ARC118925 (1 μM) on the maximal response to ATP (100 μM). The traces show the average responses from all cells in the field. The P2Y1 antagonists (MRS2179, 10 μM) had only a small effect on the response. The responses are almost fully blocked by the P2Y2 receptor blocker (ARC118925; 1 μM). The small residual response is inhibited by the P2Y1 antagonist. At the end of the experiment CCh (100 μM) was applied to confirm cell viability. (B) Composite Ca2+ images showing the cells that respond to ATP (100 μM) in presence of MRS2179 or ARC118925 (red) or both. The right panel shows the cells that response to CCh (100 μM; green) at the end of the experiment. All images are from the same field of endothelium. (C) Ca2+ signals from the activated cells in B, 30s baseline and 60s activation are shown. (D) Peak Ca2+ (F/F0) response from each individual cell (circle) matched for each treatment (grey line) from a single experiment. Average response indicated by white circle and matched across each treatment by bold line. The Plotting Density (right-hand axis) indicates the distribution of Peak F/F0 values (red high, blue low occurrence of the same values). (E) Paired summary data illustrating changes in Peak F/F0 response values averaged across all cells. Each color represents a different animal and is maintained across the different treatments (n=5). Scale bars, 50μm. p < 0.05.

Article Snippet: The arteries then were incubated with a rabbit anti-P2Y2 receptor antibody (1:100, sc-20124, Santa Cruz, Dallas, TX, USA) and CD31 (1:200, AF3628, R&D Systems,) in PBS containing 1% BSA and 2% Donkey Serum for 12 hours at 4°C.

Techniques: Activation Assay

Journal: Science signaling

Article Title: Spatially-structured cell populations process multiple sensory signals in parallel in intact vascular endothelium

doi: 10.1126/scisignal.aar4411

Figure Lengend Snippet: (A) Fluorescence localization of purinergic P2Y2 receptors and muscarinic M3 receptors in the endothelium. Representative images (from left) show the endothelial cell boundaries as revealed by PECAM-1 labelling (green; anti- CD31/PECAM-1), P2Y2 receptor (red; anti-P2Y2) distribution, M3 receptor distribution (green; fluorescent M3 receptor antagonist, 100 nM) and overlay of all three. The receptors distribution was not uniform across the endothelium and there was relatively little overlap of purinergic and muscarinic receptor staining. (B) Expanded view of the endothelial images shown in A. The expanded region is shown by the red box in A left-panel. (C) Negative control. The left panel is again PECAM-1 labeled to show cell boundary positions. The negative control for P2Y2 staining was obtained by omitting the primary antibody against P2Y2. The control for M3 staining was obtained by labelling with the fluorescent M3 receptor ligand in the presence of the M3 receptor antagonist 4-DAMP (10 μM). The right panel shows an overlay of all three. (D) Summary data showing the levels of M3 staining in the presence and absence of the antagonist 4-DAMP (green), the extent of labelling in the presence and absence of the P2Y2 primary antibody (red); and the extent of overlap of specific M3 and P2Y2 staining (grey). (E) Summary data showing the percentage of cells with M3 staining (green), P2Y2 receptor staining (red) and the percentage of cells expressing both M3 and P2Y2 receptor staining (grey) (n = 3, *p < 0.05, unpaired t-test). Scale bars, 50 μm.

Article Snippet: The arteries then were incubated with a rabbit anti-P2Y2 receptor antibody (1:100, sc-20124, Santa Cruz, Dallas, TX, USA) and CD31 (1:200, AF3628, R&D Systems,) in PBS containing 1% BSA and 2% Donkey Serum for 12 hours at 4°C.

Techniques: Fluorescence, Staining, Negative Control, Labeling, Control, Expressing