Journal: Nature

Article Title: DAXX represents a new type of protein-folding enabler.

doi: 10.1038/s41586-021-03824-5

Figure Lengend Snippet: Fig. 1 | DAXX prevents protein misfolding and aggregation. a, b, Heat-induced luciferase inactivation (a, 5 nM) and aggregation (b, 200 nM) in presence or absence of GST, DAXX and HSP70–HSP40 at 200 nM (a) or at the indicated molar ratios (b). c, Aggregation of ATXN1(82Q) (50 nM) in the presence or absence of glutathione S-transferase (GST) or DAXX (200 nM each). Raw data for this and other gels are found in Supplementary Fig 1. PE, pelletable aggregates that were soluble with SDS; SN, supernatant. d–f, Fibrillization of α-Syn (70 μM) in the presence GST, HSP70–HSP40, HSPs (HSP70–HSP40–HSP104(A503S)) (200 nM each) and DAXX (100–400 nM) was assayed by ThT-binding (d), electron microscopy (e; red arrows, fibrils; blue arrows, large oligomers; scale bar, 100 nm), and sedimentation followed by dot blot for SDS-soluble and SDS-resistant (SR) aggregates and total α-Syn and by disuccinimidyl suberate (DSS) cross-linking for soluble oligomers (f). IB, immunoblot. g–i, Fibrillization of Aβ42 monomers (10 μM) in the absence or presence of DAXX (50–600 nM) (g, h), and viability of SH-SY5Y cells treated with Aβ42 pre-incubated with or without DAXX (i). An ATP-regeneration system was included with heat-shock proteins but not DAXX (all subsequent experiments with heat-shock proteins, but not experiments with DAXX, also contained an ATP-regeneration system). Data are mean or mean ± s.d. (n = 4 for i, and 3 for the rest) and are representative of three independent experiments. *P < 0.05, NS, not significant; unpaired Student’s t-test.

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology Animals and other organisms Human research participants Clinical data Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies against the following proteins/epitopes were purchased from the indicated sources: GAPDH (sc-47724,Mouse, WB, IP, IF and IHC(P)), His (sc-8036, Mouse, IP, WB, IHC(P), ELISA, IF, FCM), GST (sc-138, Mouse, IP, WB, IHC(P), ELISA, IF, FCM), p53DO1 (sc-126, Mouse, WB, IP, IF, IHC(P) and FCM), Mdm2 (sc-965, Mouse, WB, IP, IF and IHC(P)), DAXX (sc-8043, Mouse IP,WB, IHC(P), ELISA, IF, FCM), α-synuclein syn211 (sc-12767, Mouse, IP, WB, IHC(P), ELISA, IF, and FCM) (Santa Cruz Biotechnology); Flag (#14793, Rabbit, WB, IP, IHC, ChIP, IF, FCM, ELISA) and DAXX (#4533S, Rabbit, WB, IP, IHC, ChIP, IF, FCM, ELISA) (Cell Signaling Technology); p53 (PAb1620, #OP33, Mouse, WB, IP, IF; PAb240, #OP29, Mouse, WB, IP), and Mdm2 (#OP46, Mouse, WB, IP) (Calbiochem); and HA (ab137838, Rabbit, WB, ICC/IF, IP) and Luciferase (ab21176, Rabbit, ICC/IF, WB) (Abcam); βAmyloid 1-42 (#805509) (BioLegend, Mouse, WB, IHC(P), ELISA).

Techniques: Luciferase, Binding Assay, Electron Microscopy, Sedimentation, Dot Blot, Western Blot, Incubation

Journal: Nature Communications

Article Title: In vivo cation exchange in quantum dots for tumor-specific imaging

doi: 10.1038/s41467-017-00153-y

Figure Lengend Snippet: Breast tumor-specific imaging achieved by extravascular delivery of etchable ZHS-QDs. Mice bearing orthotopic MCF10CA1a human breast tumors received an intravenous injection of iRGD or PBS before an intravenous dose of ZHS-QDs. Ag-TS was given intraperitoneally. n = 4 per group. a The mice were anesthetized and imaged with a Li-Cor Pearl imager 40 min after etching. Arrows , tumors. b In situ NIR imaging of the mice after euthanasia and necropsy performed under deep anesthesia. c NIR images of collected tissues. d NIR signal per unit area in collected tissues ( left panel ) and T/Li ratio ( right panel ). B, brain; H, heart; Li, liver; S, spleen; Lu, lung; K, kidney; T, tumor. Statistics, two-way analysis of variance ( left panel ) or Student’s t -test ( right panel ); error bars , SEM; ns, not significant; * P < 0.05; *** P < 0.001. e Confocal micrographs of cultured MCF10CA1a cells treated with ZHS-QDs with or without free iRGD followed by etching with Ag-TS. Note that the QDs were internalized into the cells in the presence of iRGD, and that only the extracellular QDs were etched. Blue, Hoechst 33342; green, ZHS-QDs; scale bars , 50 μm. Insets show a magnified view of the boxed areas . f Epifluorescence micrographs of cultured PC-3 human prostate cancer cells treated with cell-penetrating QDs followed by etching with Ag-TS. PC-3 cells were incubated with CdSe/ZnS QDs coated with a cell-penetrating peptide KCDGRPARPAR. Only speckled peri-nuclear signals remain after etching. Scale bars , 50 µm. g The tumors shown in c were processed for immunofluorescence staining and subjected to confocal microscopy. Blue , DAPI; red , CD31 or α-SMA; green , ZHS-QDs; scale bars , 50 μm. The boxed area is magnified. Note that the QD signals are found in the perinuclear area of the cells

Article Snippet: Tissue sections were treated with 0.25% Triton X-100 for 10 min, washed with PBS 3 times, blocked with 1% bovine serum albumin for 1 h, and incubated with a rat anti-mouse CD31 primary antibody (Catalog number: 553370, BD Biosciences, San Jose, CA), rabbit anti-mouse α-SMA antibody (Product code: ab5694, Abcam, Cambridge, MA), or a rat anti-mouse ER-TR7 antibody (Catalog number: sc-73355, Santa Cruz Biotechnology, Dallas, TX) at 4 °C overnight.

Techniques: Imaging, Injection, In Situ, Cell Culture, Incubation, Immunofluorescence, Staining, Confocal Microscopy

Journal: Nature Communications

Article Title: In vivo cation exchange in quantum dots for tumor-specific imaging

doi: 10.1038/s41467-017-00153-y

Figure Lengend Snippet: Desmoplastic PDAC imaging with etchable ZHS-QDs. a H&E staining of orthotopic KRAS-Ink tumor tissue collected from mice. A mixed histology ranging from pancreas tissue with severe desmoplasia, high-grade PanIN, to full-blown PDAC is observed. n = 3, scale bars , 100 μm. b – e Mice bearing orthotopic KRAS-Ink PDAC tumors received intravenous injections of iRGD or PBS 25 min before ZHS-QD injection. Ag-TS was intravenously injected 30 min after the QD injection. n = 3 per group. NIR images of anesthetized mice b and collected tissues c . Arrows , tumors; dotted lines , tissues. B, brain; H, heart; Li, liver; S, spleen; Lu, lung; K, kidney; T, tumor. NIR signal per unit area in collected tissues ( left panel ) and T/Li ratio ( right panel ) d . Statistics, two-way analysis of variance ( left panel ) or Student’s t -test ( right panel ); error bars , SEM; ns, not significant; * P < 0.05; *** P < 0.001. Confocal micrographs of tumor sections e . Blue , DAPI; red , CD31, ER-TR7 or α-SMA; green , ZHS-QDs; scale bars , 50 μm; P stands for PanIN. Inset , an area with PanINs

Article Snippet: Tissue sections were treated with 0.25% Triton X-100 for 10 min, washed with PBS 3 times, blocked with 1% bovine serum albumin for 1 h, and incubated with a rat anti-mouse CD31 primary antibody (Catalog number: 553370, BD Biosciences, San Jose, CA), rabbit anti-mouse α-SMA antibody (Product code: ab5694, Abcam, Cambridge, MA), or a rat anti-mouse ER-TR7 antibody (Catalog number: sc-73355, Santa Cruz Biotechnology, Dallas, TX) at 4 °C overnight.

Techniques: Imaging, Staining, Injection