Journal: eLife

Article Title: VEGF-C promotes the development of lymphatics in bone and bone loss

doi: 10.7554/eLife.34323

Figure Lengend Snippet:

Article Snippet: Plasma was collected from mice and commercially available ELISA kits were used to measure the circulating levels of VEGF-C (CUSABIO, #CSB-E07361M, Houston, TX) and CTX-1 (CUSABIO, #CSB-E12782M).

Techniques: Blocking Assay, Enzyme-linked Immunosorbent Assay, Isolation, cDNA Synthesis, Software

Journal: Cell Death & Disease

Article Title: Colon cancer cell treatment with rose bengal generates a protective immune response via immunogenic cell death

doi: 10.1038/cddis.2016.473

Figure Lengend Snippet: RB treatment induced the release and/or increased expression of DAMPs in colon cancer cells. ( a and b ) RB treatment of CT26 colon cancer cells induced increased surface expression of CRT in a dose-dependent manner ( a ). Increased expression was observed on both dead ( b , upper panel) and live cells ( b , lower panel) based on flow cytometry analysis after 1 h of treatment. ( a ) A representative example of three separate experiments summarized in ( b ). ( c ) CRT expression increased minutes after exposure to RB, reaching significance by 10 min. ( d and e ) ATP content in colon cancer cells was determined by cytofluorometric detection of intracellular ATP with quinacrine, expressed as the percentage of MFI relative to untreated cells. A representative histogram ( d ) and summary of three separate experiments ( e ) demonstrate a decrease in intracellular ATP levels upon treatment with RB. ( f ) HMGB1 secretion significantly increased as measured in culture supernatants after treatment with 200 μ M RB for 30 min. ( g and h ) RB treatment resulted in increased membrane expression and reduced cytosolic levels of HSP90 as the time of treatment increased from 30 min to 6 h as measured on western blot analysis ( g ) and after densitometric correction for housekeeping gene loading controls ( h ) (* P <0.05 and ** P <0.01)

Article Snippet: HMGB1 levels were determined using a Mouse HMGB1 ELISA Kit from Novatein Biosciences (Woburn, MA, USA) according to the protocol.

Techniques: Expressing, Flow Cytometry, Membrane, Western Blot

Journal: eLife

Article Title: Csf1 from marrow adipogenic precursors is required for osteoclast formation and hematopoiesis in bone

doi: 10.7554/eLife.82112

Figure Lengend Snippet: ( A ) Representative fluorescent TRAP staining images of femoral long bones from control and Csf1 CKO Adipoq mice at 3 months of age show TRAP+ osteoclasts at different skeletal sites: secondary spongiosa (SS), chondro-osseous junction (COJ), and endosteal surface (Endo.S). TB: trabecular bone; CB: cortical bone. Scale bar = 50 μm. ( B ) Quantification of osteoclast surface (Oc.S) at three skeletal sites. BS: bone surface. L: COJ length. n=5 mice/group. ***, p<0.001 CKO vs control. ( C ) Representative TRAP staining images of osteoclast culture derived from control and Csf1 CKO Adipoq BMMs at 7 days after addition of RANKL and Csf1. Arrows point to mature osteoclasts. Scale bar = 200 μm. ( D ) Quantification of TRAP+ multinucleated cells (>3 nuclei/cell) per field. n=7 mice/group. ( E ) Representative Osterix staining of trabecular bone from control and Csf1 CKO Adipoq femurs. Scale bar = 50 μm. ( F ) Quantification of osteoblast surface (OB.S). BS, bone surface. n=8–12 mice/group. ( G ) Representative double labeling of trabecular bone from control and Csf1 CKO Adipoq femurs. ( H ) Bone formation activity is quantified. MAR: mineral apposition rate; MS: mineralizing surface; BFR: bone formation rate. n=4 mice/group. ( I ) Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (PINP) in control and CKO mice. n=6–8 mice/group. *, p<0.05 CKO vs control. Figure 4—source data 1. Full dataset for . Figure 4—source data 2. Full dataset for . Figure 4—source data 3. Full dataset for . Figure 4—source data 4. Full dataset for . Figure 4—source data 5. Full dataset for .

Article Snippet: Sera were collected during mouse euthanization for measuring bone turnover markers, collagen type I C-telopeptide degradation products (mouse CTX-I ELISA Kit, AFG Scientific, Northbrook, IL, USA) and N-terminal propeptide of type I procollagen (Immunotag Mouse PINP ELISA Kit, G-Bioscience, St. Louis, MO, USA) respectively, according to the manufacturer’s instructions.

Techniques: Staining, Control, Derivative Assay, Labeling, Activity Assay, Enzyme-linked Immunosorbent Assay, Marker