Journal: Cancer research

Article Title: Mobilizing transit-amplifying cell-derived ectopic progenitors prevents hair loss from chemotherapy or radiation therapy

doi: 10.1158/0008-5472.CAN-17-0667

Figure Lengend Snippet: Remodeling of ORS cells for the late regenerative attempt after 5.5Gy of IR. A, Lineage tracing in K5CreER; R26LSLtdTomato mice. B, Cell proliferation mapped by pulse BrdU labeling. The p-cadherin+ lower tip cells started to proliferate at 84hrs (white arrowheads) and regenerated new hair bulb at 96hrs. C, Analysis of BgSC apoptosis by TUNEL. D, Continuous BrdU labeling showed that BgSCs did not proliferate until day 5 (white arrowheads). E, Lineage tracing of BgSCs in K19CreER; R26LSLtdTomato mice. Labeled BgSC progeny did not move out of the bulge until day 5. F, Immunofluorescence of p-cadherin and Sox9. Similar to SHG (yellow arrowheads), p-cadherin+ lower tip cells were also positive for Sox9 (white arrowheads). G, Lgr5 promoter-driven EGFP expression in Lgr5EGFP-Ires-CreERT2 mice. The lower tip cells were positive for EGFP. H, Expression of ShgSC specific genes. Similar to activated ShgSCs, several specific genes were upregulated in lower tip cells at 72hrs. I, Possible cell origin for late regenerative attempts. J, Lineage tracing in Lgr5EGFP-Ires-CreERT2; R26LSLtdTomato mice. Error bars represent mean ± S.E.M. Scale bar=25µm.

Article Snippet: The following antibodies were used: CD34-FITC (eBioscience, 11-0341, 1:50), Sca1-PE-Cy7 (eBioscience, 25-5981, 1:50) and p-cadherin-PE (R&D systems, FAB761P, 1:100).

Techniques: Labeling, TUNEL Assay, Immunofluorescence, Expressing

Journal: eLife

Article Title: Concerted IL-25R and IL-4Rα signaling drive innate type 2 effector immunity for optimal helminth expulsion

doi: 10.7554/eLife.38269

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-Arginase-1 (Polyclonal) , R and D Systems , IC5868P.

Techniques: Recombinant, Staining, Control

Journal: bioRxiv

Article Title: CD36-Pyruvate Kinase M2 Signaling Promotes Macrophage Phagocytosis Through Mitochondrial Reactive Oxygen Species

doi: 10.1101/2023.09.07.556574

Figure Lengend Snippet: A , a diagram of the ex vivo mtROS and phagocytosis assay. B , Total amount of aortic F4/80 + /Trem2 + macrophages were quantified and shown in the bar graph; n=4-7 individual mice per group. C , MitoNeoD MFI was quantified in aortic F4/80 + /Trem2 + macrophages and shown in the bar graph; n=5 individual mice per group. D , pHrodo MFI was quantified in aortic F4/80 + /Trem2 + macrophages and shown in the bar graph; n=5-6 individual mice per group. E , a diagram of the in vivo phagocytosis assay. F , pHrodo MFI was quantified in aortic F4/80 + /Trem2 + macrophages and shown in the bar graph; n=5-7 individual mice per group.

Article Snippet: For phagocytosis assays, the filtered single-cell suspension was incubated with 20 μg/ml pHrodo-conjugated E. coli bioparticles in RPMI1640 with 10% FBS at 37°C for 30 min, followed by immunostaining with PE/Cy5-conjugated F4/80 antibody (BioLegend, Cat#123111) and PE-conjugated Trem2 antibody (R&D Systems, Cat#FAB17291P) in Flow Buffer at RT for 15 min in the dark.

Techniques: Ex Vivo, Phagocytosis Assay, In Vivo

Journal: bioRxiv

Article Title: CD36-Pyruvate Kinase M2 Signaling Promotes Macrophage Phagocytosis Through Mitochondrial Reactive Oxygen Species

doi: 10.1101/2023.09.07.556574

Figure Lengend Snippet: A-D , Mouse scRNA-seq data were re-analyzed from a previous publication . Uniform manifold approximation and projection (UMAP) representation of 11 aortic CD45 + immune cell clusters were shown in A . Trem2 gene expression pattern ( B ) and Pkm gene expression pattern ( C ) were shown in the UMAP. D , Violin plots show the Pkm and Trem2 expression distribution among aortic macrophage subpopulations. ResMac: resident macrophages; InflaMac: inflammatory macrophages. E , HMDMs transfected with PKM siRNA were treated with 50 μg/ml oxLDL for 24 h before subjected to mtROS assay (left panel) or phagocytosis assay (right panel). MFI was quantified and shown in the bar graph; n=4-5 per group. F , Representative confocal images of macrophages immunostained for PKM2 (green) and Tom20 (red). Nuclei were stained by DAPI (blue). Scale bar: 5 μm. G , HMDMs treated with 20 μg/ml LDL (control) or oxLDL for 3 h were lysed, subjected to cell fractionation into mitochondrial and cytosol fractions. PKM2 and ATP5A (mitochondria fraction loading control) blot images from mitochondrial fractions were shown on the left. PKM2 and β-actin (cytosol fraction loading control) blot images from cytosol fractions were shown on the right. Images were quantified, normalized to each loading control, and expressed as fold change of control. n=4 per group. H , WT or Cd36 -null peritoneal macrophages treated with 20 μg/ml LDL (control) or oxLDL for 3 h and then processed as in G. Mitochondrial fractions were immunoblotted for PKM2 and ATP5A and blot images were shown. Images were quantified, normalized, and expressed as fold change of control. n=4 per group. I , WT macrophages treated with 20 μg/ml oxLDL or pre-treated 1 or 5μM shikonin before addition of oxLDL, incubating for 3 h, and then processed as in G. Mitochondrial fractions were immunoblotted for PKM2 and Tom20 and blot images were shown. Images were quantified and expressed as fold change of control. n=3 per group. J , WT macrophages pre-treated with 20 μg/ml oxLDL or in combination with 1 μM shikonin for 24 h before mtROS or phagocytosis assay. The MitoNeoD (left) or pHrodo (right) MFI was quantified and shown in the bar graph; n=3-4 per group.

Article Snippet: For phagocytosis assays, the filtered single-cell suspension was incubated with 20 μg/ml pHrodo-conjugated E. coli bioparticles in RPMI1640 with 10% FBS at 37°C for 30 min, followed by immunostaining with PE/Cy5-conjugated F4/80 antibody (BioLegend, Cat#123111) and PE-conjugated Trem2 antibody (R&D Systems, Cat#FAB17291P) in Flow Buffer at RT for 15 min in the dark.

Techniques: Gene Expression, Expressing, Transfection, Phagocytosis Assay, Staining, Control, Cell Fractionation

Journal: Cell reports

Article Title: Expeditious recruitment of circulating memory CD8 T cells to the liver facilitates control of malaria

doi: 10.1016/j.celrep.2021.109956

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD11b-PE (Clone M1/70) , Tonbo , Cat# 50-0112-U100.

Techniques: Control, Blocking Assay, Virus, Luciferase, Expressing, Recombinant, Staining, Cell Isolation, RNA Sequencing, Software