Journal: Theranostics

Article Title: In Vivo Targeting of Metabolically Labeled Cancers with Ultra-Small Silica Nanoconjugates

doi: 10.7150/thno.16003

Figure Lengend Snippet: (a) Schematic illustration of Ac 4 ManDBCO-mediated metabolic labeling of cancer cells and subsequent detection of cell-surface DBCO groups using Cy5-azide via copper-free Click chemistry. (b) CLSM images of LS174T cells after treatment with Ac 4 ManDBCO (20 μM) or PBS for three days and further incubation with Cy5-azide (20 μM) for 30 min. Cell nuclei were stained with DAPI (blue). Scale bar represents 10 μm. (c) Flow cytometry analyses of LS174T cells following the same treatment in (b). (d) Mean Cy5 fluorescence intensity of LS174T cells extracted from (b). Data were presented as mean ± SEM (n=3) and analyzed by Student's t-test (two-tailed) (0.01 < *P ≤ 0.05, and 0.001 < **P ≤ 0.01).

Article Snippet: Cy5-azide was purchased from Kerafast (Boston, MA, USA).

Techniques: Labeling, Incubation, Staining, Flow Cytometry, Fluorescence, Two Tailed Test

Journal: Theranostics

Article Title: In Vivo Targeting of Metabolically Labeled Cancers with Ultra-Small Silica Nanoconjugates

doi: 10.7150/thno.16003

Figure Lengend Snippet: (a) Schematic illustration of the preparation of azido-/Cy5-NCs and 64 Cu-/azido-/Cy5-NCs. The chemical structures of TEOS, DOTA-silane, Cy5-silane, and azido-PEG 3.4k -silane were shown. Cy5-NCs and 64 Cu-/Cy5-NCs were synthesized similarly except that azido-PEG 3.4k -silane was replaced with mPEG 3.4k -silane. (b) SEM image of azido-/Cy5-NCs showed a size of 48.8 ± 4.9 nm. Scale bar represents 200 nm. (c) Fluorescence spectra of azido-/Cy5-NCs and Cy5-NCs. The excitation wavelength was set at 650 nm. (d) FTIR spectra of silica core, azido-/Cy5-NCs and Cy5-NCs. Inset: enlarged view of transmittance profiles of azido-/Cy5-NCs and Cy5-NCs within the wavenumber range of 2095-2114 cm -1 . (e) Loss of 64 Cu over time from 64 Cu-/azido-/Cy5-NCs or 64 Cu-/Cy5-NCs following incubation at 37 o C in 10% FBS (n=3).

Article Snippet: Cy5-azide was purchased from Kerafast (Boston, MA, USA).

Techniques: Synthesized, Fluorescence, Incubation

Journal: Theranostics

Article Title: In Vivo Targeting of Metabolically Labeled Cancers with Ultra-Small Silica Nanoconjugates

doi: 10.7150/thno.16003

Figure Lengend Snippet: (a) Schematic illustration of the targeted and non-targeted internalization of azido-/Cy5-NCs by cancer cells. (b) Flow cytometry profiles of LS174T cells following treatment with either Ac 4 ManDBCO (blue curve) or PBS (red curve) for 72 h and further incubation with azido-/Cy5-NCs for 30 min. Cells without azido-/Cy5-NCs treatment served as negative controls (black curve). (c) Mean fluorescence intensity of LS174T cells after incubation with azido-/Cy5-NCs or Cy5-NCs for 0.5, 1, and 2 h, respectively following pre-treatment with Ac 4 ManDBCO for 72 h. Cells treated with azido-/Cy5-NCs following pretreatment with PBS served as controls (n=6). Statistically significantly lower Cy5 fluorescence intensity than Ac 4 ManDBCO+azido-/Cy5-NCs group was observed in both PBS+azido-/Cy5-NCs and Ac 4 ManDBCO+Cy5-NCs groups (0.01 < * P ≤ 0.05, and 0.001 < ** P ≤ 0.01).

Article Snippet: Cy5-azide was purchased from Kerafast (Boston, MA, USA).

Techniques: Flow Cytometry, Incubation, Fluorescence

Journal: Theranostics

Article Title: In Vivo Targeting of Metabolically Labeled Cancers with Ultra-Small Silica Nanoconjugates

doi: 10.7150/thno.16003

Figure Lengend Snippet: (a) Time frame of in vivo imaging study. LS174T tumors were implanted in both flanks of athymic nude mice. When the tumors reached ~50 mm 3 , Ac 4 ManDBCO (5 mg/kg) was intratumorally injected into the left tumors once daily for three days (Day 1-3), while the right tumors were injected with PBS as controls. On Day 4, 64 Cu-/azido-/Cy5-NCs or 64 Cu-/Cy5-NCs (~100 μCi) were i.v. injected and their biodistribution were monitored via PET/CT imaging. (b) PET/CT imaging of athymic nude mouse at 24 h post injection of 64 Cu-/azido-/Cy5-NCs. The tumor area was zoomed in to clearly show the retention of 64 Cu-/azido-/Cy5-NCs in the left and right tumors. Tumors were shown by the arrows. Inset: calculated radioactivity of the left and right tumors using the imaging software. (c) Biodistribution of 64 Cu-/azido-/Cy5-NCs and 64 Cu-/Cy5-NCs at 24 h post injection, as determined by ex vivo radioactivity measurement on a λ-counter. Data were presented as mean ± SEM (n=3) and analyzed by Student's t-test (two-tailed) (0.001 < **P ≤ 0.01).

Article Snippet: Cy5-azide was purchased from Kerafast (Boston, MA, USA).

Techniques: In Vivo Imaging, Injection, Positron Emission Tomography-Computed Tomography, Imaging, Radioactivity, Software, Ex Vivo, Two Tailed Test

Journal: Theranostics

Article Title: In Vivo Targeting of Metabolically Labeled Cancers with Ultra-Small Silica Nanoconjugates

doi: 10.7150/thno.16003

Figure Lengend Snippet: (a) Whole-body fluorescence imaging of athymic nude mice bearing LS174T tumors at 1, 6, 24, 48, and 72 h post injection of azido-/Cy5-NCs. Tumors were shown by the arrows. The left tumors were injected with Ac 4 ManDBCO (5 mg/kg) once daily for three days (Day 1-3), while the right tumors were injected with PBS as controls. Azido-/Cy5-NCs or Cy5-NCs were i.v. injected on Day 4. (b) Ex vivo imaging of tissues harvested at 72 h post injection of azido-/Cy5-NCs. (c) Mean Cy5 fluorescence intensity of tissues extracted from ex vivo images (n=3). (d) Representative CLSM images of tissue sections of the left and right tumors from mice treated with azido-/Cy5-NCs. Cell nuclei were stained with DAPI (blue).

Article Snippet: Cy5-azide was purchased from Kerafast (Boston, MA, USA).

Techniques: Fluorescence, Imaging, Injection, Ex Vivo, Staining

Journal: International Journal of Molecular Sciences

Article Title: Interplay between Inter-Subunit Rotation of the Ribosome and Binding of Translational GTPases

doi: 10.3390/ijms24086878

Figure Lengend Snippet: LepA stabilizes the non-rotated conformation of the ribosome. Histograms show FRET distribution in S6-Cy5/L9-Cy3 ribosomes. Ribosomes bound with P-site deacylated tRNA fMet ( a – d ) were incubated with RF3•GDPNP ( b ) or LepA•GDPNP (c) or LepA•GDPCP ( d ). Ribosomes bound with P-site deacylated tRNA Phe and A-site N-Ac-Tyr-tRNA Tyr ( e , f ) were incubated with LepA•GDPNP ( f ). N indicates the total number of FRET traces compiled into each histogram. % fluc designates the percentage of traces that show fluctuations. Yellow lines show individual Gaussian fits. Black line indicates the sum of Gaussian fits. The fraction of the ribosomes in the R and NR conformations are shown above the corresponding 0.37 and 0.56 Gaussian peaks.

Article Snippet: Cy3 and Cy5 maleimide were purchased from Click Chemistry Tools; GDPNP—Jena Biosciences (Jena, Germany); GDPCP—Sigma Aldrich (St. Louis, MI, USA).

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: Interplay between Inter-Subunit Rotation of the Ribosome and Binding of Translational GTPases

doi: 10.3390/ijms24086878

Figure Lengend Snippet: EF-G stabilizes the rotated conformation of the ribosome. Histograms show FRET distribution in S6-Cy5/L9-Cy3 ribosomes. Ribosomes bound with P-site deacylated tRNA fMet ( a , b ) were incubated with either EF-G•GDPNP ( a ) or EF-G•GDPCP ( b ). Ribosomes bound with P-site acylated N-Ac-Tyr-tRNA Tyr ( c , d ) were incubated with EF-G•GDPNP ( d ). N indicates the total number of FRET traces compiled into each histogram. % fluc designates the percentage of traces that show fluctuations. Yellow lines show individual Gaussian fits. Black line indicates the sum of Gaussian fits. The fraction of the ribosomes in the R and NR conformations are shown above the corresponding 0.37 and 0.56 Gaussian peaks.

Article Snippet: Cy3 and Cy5 maleimide were purchased from Click Chemistry Tools; GDPNP—Jena Biosciences (Jena, Germany); GDPCP—Sigma Aldrich (St. Louis, MI, USA).

Techniques: Incubation