Journal: Pharmaceuticals

Article Title: A Short-Term Biological Indicator for Long-Term Kidney Damage after Radionuclide Therapy in Mice

doi: 10.3390/ph10020057

Figure Lengend Snippet: Incidence of γ-H2AX-positive nuclei in the renal cortex of mice euthanized two weeks after the injection of ( A ) saline, ( B ) 10 MBq, ( C ) 20 MBq and ( D ) 30 MBq 177 Lu-folate, or three months after injection of ( E ) saline, ( F ) 10 MBq, ( G ) 20 MBq, and ( H ) 30 MBq 177 Lu-folate. Positive nuclei typically exhibited one to multiple intensely brown, round foci of approximately 1 µm in diameter, or a more diffuse light brown staining. The number of γ-H2AX-positive nuclei is markedly increased in a dose-related manner in mice at two weeks ( B – D ), and to a lesser extent and only in the mid- and high-dose groups in mice at three months ( G , H ). γ-H2AX-positive nuclei in tubular epithelial cells, glomeruli and interstitium are indicated with arrows, open arrowheads and solid arrowheads, respectively.

Article Snippet: This was followed by incubation with rabbit anti-mouse γ-H2AX antibody (polyclonal antibody #2577, Cell Signaling Technology, Danvers, MA, USA, 1:50 dilution in Dako antibody diluent, Dako-Agilent Technologies, Denmark) overnight at 4 °C.

Techniques: Injection, Staining

Journal: Pharmaceuticals

Article Title: A Short-Term Biological Indicator for Long-Term Kidney Damage after Radionuclide Therapy in Mice

doi: 10.3390/ph10020057

Figure Lengend Snippet: ( A ) Percentage of γ-H2AX-positive nuclei and ( B ) percentage of positive nuclei per glomerulus (N/G) in kidneys of mice two weeks after injection of 177 Lu-folate. Compared to the kidneys of untreated mice which showed rare positive cells, there is a dose-related, statistically significant increase in the number of γ-H2AX-positive nuclei in mice euthanized two weeks after injection (≥10 MBq 177 Lu-folate). The highest activity of 177 Lu-folate was associated with a statistically significant increase in the number of γ-H2AX-positive nuclei in glomerular cells at the two-week time point (B). * p < 0.05.

Article Snippet: This was followed by incubation with rabbit anti-mouse γ-H2AX antibody (polyclonal antibody #2577, Cell Signaling Technology, Danvers, MA, USA, 1:50 dilution in Dako antibody diluent, Dako-Agilent Technologies, Denmark) overnight at 4 °C.

Techniques: Injection, Activity Assay

Journal: Pharmaceuticals

Article Title: A Short-Term Biological Indicator for Long-Term Kidney Damage after Radionuclide Therapy in Mice

doi: 10.3390/ph10020057

Figure Lengend Snippet: Positive correlation between the number of γ-H2AX-positive nuclei in the kidneys of mice treated with different activities of 177 Lu-folate (10 MBq, 20 MBq and 30 MBq) and untreated control mice after two weeks and long-term final scores of histopathological changes. Dose-related increases in the number of γ-H2AX-positive nuclei, as a biological indicator of radiation effects, correlated well with long-term kidney damage.

Article Snippet: This was followed by incubation with rabbit anti-mouse γ-H2AX antibody (polyclonal antibody #2577, Cell Signaling Technology, Danvers, MA, USA, 1:50 dilution in Dako antibody diluent, Dako-Agilent Technologies, Denmark) overnight at 4 °C.

Techniques:

Journal: Molecular Cancer

Article Title: The role of TGFBI (βig-H3) in gastrointestinal tract tumorigenesis

doi: 10.1186/s12943-015-0335-z

Figure Lengend Snippet: TGFBI promotes liver cell survival and proliferation after irradiation. (A) Aml-12 liver cells overexpressed TGFBI after stable transfection with the TGFBI-expressing construct pCEP4-TGFBI. TGFBI mRNA levels of transfected Aml-12 liver cells were measured by RT-qPCR. β-actin mRNA levels served as internal controls, and data are expressed as ratios of TGFBI versus β-actin signals (left panel). The supernatants of these cells were quantified by ELISA in duplicate for TGFBI protein levels; regular culture medium was included as an additional control (right panel). Means ± SD of representative experiments are shown. (B) TGFBI promotes post-irradiation survival of liver cells carrying DNA damage. TGFBI-overexpressing Aml-12 and control cells were irradiated at 20 Gy and cultured for 20 h. They were then stained with Abs against γH2AX and caspase-3, and analyzed by flow cytometry. The percentages of caspase-3-positive cells among γH2AX-positive cells are presented (left), and the percentages of 3 independent experiments are summarized in the bar graph (right), with p -value indicated (2-tailed Student’s t test). (C) TGFBI promotes post-irradiation proliferation of liver cells. TGFBI-overexpressing Aml-12 and control cells after irradiation were cultured in complete DMEM/F12 medium containing 10% FCS for 40 h. 3 H-thymidine was added 16 h before the cells were harvested. Thymidine uptake of both irradiated and non-irradiated cells in a representative experiment is shown on the left. Post-irradiation proliferation indices of 3 independent experiments are summarized in a bar graph on the right with p -value indicated (2-tailed Student’s t test).

Article Snippet: They were first fixed and permeabilized using the eBioscience Fixation/Permeabilization Kit (eBioscience, San Diego, CA), and then stained with PE-conjugated rabbit anti-mouse phospho-histone H2AX (γH2AX) Ab and Alexa 488-conjugated rabbit anti-mouse cleaved caspase-3 Ab (Cell Signaling Technology).

Techniques: Irradiation, Stable Transfection, Expressing, Construct, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Staining, Flow Cytometry