Journal: Journal of Translational Medicine

Article Title: Chimeric antigen receptor T cells targeting cell surface GRP78 efficiently kill glioblastoma and cancer stem cells

doi: 10.1186/s12967-023-04330-0

Figure Lengend Snippet: Pep42-BBZ CAR-T cells showed antitumor activity in vivo. A Schematic in the top panel illustrates the experiment design for the U-251MG xenograft tumors. NCG mice was inoculated subcutaneously with 5 × 10 6 luciferase-expressing U-251MG cells. Five days later mock-BBZ and Pep42-BBZ CAR-T cells (1 × 10 7 per mouse) were injected intravenously into tail veins. The tumors growth was monitored once a week. B A representative image of csGRP78 staining of xenograft tumor sections by IF. Five days after tumor cells inoculation, tumor tissues were extracted and cell surface IF was performed for GRP78. Control was stained without GRP78 antibody. C The tumor bioluminescence images by IVIS imaging. D Tumor bioluminescence intensity curves for the two groups. E Representative CD3 staining of tumor sections by IF. Five-days post injection, 3 tumors from each group were extracted, embedded in paraffin, sectioned, and stained with CD3ζ antibody coupled with Cy3-conjunctated secondary antibodies. Two representative images are included from each group. F The statistics and representatives of CD4 and CD8 staining on Pep42-BBZ CAR-T treated tumor sample sections. The numbers of FITC labeled-CD4 and Cy3 labeled-CD8 were calculated and performed for percentage counting. G Representative NESTIN staining of tumor sections by IF. Five-days post injection, 3 tumors from each group were extracted, embedded in paraffin, sectioned, and stained with NESTIN antibodies coupled with Cy3-conjunctated secondary antibodies. Two representative images from each group are shown. Scale bar = 100 μm

Article Snippet: Tumor growth was monitored once a week using an in vivo imaging software (IVIS) system (Lumina Xr, PerkinElmer, USA).

Techniques: Activity Assay, In Vivo, Luciferase, Expressing, Injection, Staining, Control, Imaging, Labeling

Journal: Nature Protocols

Article Title: A surgical orthotopic approach for studying the invasive progression of human bladder cancer

doi: 10.1038/s41596-018-0112-8

Figure Lengend Snippet: Fig. 3 | End-stage bladder cancer xenograft growth as observed with different methods. a–c, UM-UC-9 (a), UM-UC-6 (b) and SW 780 (c) bladder cancer xenografts in NOD-SCID mice are shown as examples. a, In vivo bioluminescence imaging with the IVIS Spectrum Optical Imaging System, displaying the absence (left panel) or presence (right panel) of tumor at the anatomical bladder localization. Scale bar indicates radiance (photons). b, Similar experiment with quantum dot–labeled cancer cells. The tumor presence was evaluated by epifluorescence via the IVIS system. These composite images are the result of spectral unmixing, which was used to distinguish between quantum dot (Qtracker 655) fluorescence and the autofluorescence of the food. The autofluorescence caused by the food in the intestines is red, whereas the tumor-specific signal is yellow. c, Representative anatomic and histological observations at the experimental end point. Top left, a bladder before excision from the mouse abdominopelvic cavity. The tumor is easily observed within the bladder. Scale bar, 5 mm. Top right, ex vivo fluorescence scanning by the IVIS system, showing tumor growth within the bladder lumen (right panel). Yellow areas indicate high fluorescence signal originating from the cancer cells labeled with quantum dots (Qtracker 525). No signal is observed in the control bladder (left panel). Scale bars, 5 mm. Red–yellow scale indicates epifluorescence units. Bottom left, excised bladders can be photographed using a dissecting stereomicroscope to record gross anatomopathological changes. Bottom right, representative 5-µm tissue section of a bladder tumor xenograft stained with H&E. Control images represent mice that underwent control sham surgeries. Scale bars for bottom images, 1 mm. The procedures were approved by the Institutional Animal Care and Use Committee of the University of Michigan and followed the animal welfare recommendations of the National Institutes of Health.

Article Snippet: Software and code Policy information about availability of computer code Data collection Living Image Software for IVIS Spectrum Imaging Systems, PerkinElmer.

Techniques: In Vivo, Imaging, Optical Imaging, Labeling, Ex Vivo, Control, Staining