Journal: STAR Protocols

Article Title: Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue

doi: 10.1016/j.xpro.2021.100971

Figure Lengend Snippet:

Article Snippet: Cy-3B Mono NHS-ester , Cytiva , PA63101.

Techniques: Recombinant, Saline, Electron Microscopy, Plasmid Preparation, Software, Imaging, Control, Mass Measurement, Capsules, Microscopy, Staining, Adhesive, Spectrophotometry

Journal: Journal of Clinical Investigation

Article Title: Mycobacterium tuberculosis hijacks host TRIM21- and NCOA4-dependent ferritinophagy to enhance intracellular growth

doi: 10.1172/jci159941

Figure Lengend Snippet: Figure 6. Ncoa4 deficiency in myeloid cells increases host resistance to Mtb. (A) Immunoblot analysis of FTH1 in BMDMs from Ncoa4+/+ and Ncoa4–/– mice infected with H37Rv for 24 hours. (B) In vitro CFUs in BMDMs from Ncoa4+/+ and Ncoa4–/– mice. (C) Representative images of H37Rv-infected BMDMs stained with FeRhoNox and LAMP-1 (scale bars: 7 μm). The colocalization ratio was quantified from 30 macrophages (right). (D) Detection of free iron in BMDMs by FeRhoNox after H37Rv infection for 72 hours. (E) The fold changes of iron-response genes in H37Rv isolated from the indicated BMDMs. (F–I) Ncoa4+/+ and Ncoa4–/– mice were aerosol-infected with about 200 CFU per mouse of H37Rv, and lungs were removed at 4 and 8 weeks postinfection (wpi). The burden of H37Rv in the lungs of Ncoa4+/+ and Ncoa4–/– mice (4 wpi, n = 5 per group; 8 wpi, n = 7 per group) was assessed by counting of CFUs (F) and acid-fast staining (G, 4 wpi; scale bars: 100 μm [top] and 20 μm [bottom]. Histopathology was assessed in lung sections stained with H&E and quantified by immune cell infiltration ratio (H, 4 wpi; scale bars: 2 mm [top] and 100 μm [bottom]). FTH1 (green) in macrophages (red, CD68-positive) was immuno- fluorescently stained (scale bars: 20 μm) and quantified by percentage (I; n = 100). Data in A–D and F are representative of 2 or 3 independent experiments. Data are presented as means ± SD; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by Student’s unpaired t test (C, H, and I) or Student’s 2-tailed unpaired t test (B, D, and F).

Article Snippet: The macrophages were washed twice with PBS and stained with 500 μL 0.5 μM Calcein-AM (BioLegend) or 5 μM FeRhoNox (Goryo Chemical) in PBS for 30 minutes at 37°C.

Techniques: Western Blot, Infection, In Vitro, Staining, Isolation, Aerosol, Histopathology