“…Scale bars show 1,000 μm (left) and 100 μm (right). Figure reprinted with permission from Yu et al., 2021 . …”
Section: Expected Outcomesmentioning
confidence: 99%
“… Figure 4 Analysis of CTL activity in tumors that are sensitive to anti-PD-1 therapy by flow cytometry Representative images of flow cytometry analysis of CD8 + , GZMB + , and IFN-γ + Cells in TDCL tumors, CD45 + cells were isolated from subcutaneous primary tumor tissues (A), plots showing the mean percentage ± SEM of CD8 + T cells in CD45 + cells, GZMB + cells, and IFN-γ + cells in tumor treated with IgG and aPD-1(n=6 for each group), two-tailed t test (B). Figure reprinted with permission from Yu et al., 2021 . Figure 5 Analysis of CD8 + T cell infiltration by immunofluorescence (A) Frozen sections of tumor tissue were stained for CD8 (green) and DAPI (blue).…”
Section: Expected Outcomesmentioning
confidence: 99%
“…Data are mean ± SEM, two tailed T test. Figure reprinted with permission from Yu et al., 2021 . Figure 6 Lentivirus-sucrose cocktail in ultracentrifuge tube Related to step 2b.…”
Section: Expected Outcomesmentioning
confidence: 99%
“… Representative images of flow cytometry analysis of CD8 + , GZMB + , and IFN-γ + Cells in TDCL tumors, CD45 + cells were isolated from subcutaneous primary tumor tissues (A), plots showing the mean percentage ± SEM of CD8 + T cells in CD45 + cells, GZMB + cells, and IFN-γ + cells in tumor treated with IgG and aPD-1(n=6 for each group), two-tailed t test (B). Figure reprinted with permission from Yu et al., 2021 . …”
Section: Expected Outcomesmentioning
confidence: 99%
“…This protocol can also be applied to other cancer types and immunotherapies. For complete details on the use and execution of this protocol, please refer to Yu et al. (2021) …”
Summary
Anti-PD-1/PD-L1 therapy shows long-term effects in many cancer types, but resistance and relapse remain the main limitations of this therapy. Here, we describe a protocol to evaluate the tumor response to immunotherapy in a mouse lung cancer model. The protocol includes the establishment of the lung cancer mouse model, anti-PD-1 treatment, tumor-infiltrating lymphocyte isolation, immunofluorescence, and flow cytometry analysis. This protocol can also be applied to other cancer types and immunotherapies.
For complete details on the use and execution of this protocol, please refer to
Yu et al. (2021)
“…Scale bars show 1,000 μm (left) and 100 μm (right). Figure reprinted with permission from Yu et al., 2021 . …”
Section: Expected Outcomesmentioning
confidence: 99%
“… Figure 4 Analysis of CTL activity in tumors that are sensitive to anti-PD-1 therapy by flow cytometry Representative images of flow cytometry analysis of CD8 + , GZMB + , and IFN-γ + Cells in TDCL tumors, CD45 + cells were isolated from subcutaneous primary tumor tissues (A), plots showing the mean percentage ± SEM of CD8 + T cells in CD45 + cells, GZMB + cells, and IFN-γ + cells in tumor treated with IgG and aPD-1(n=6 for each group), two-tailed t test (B). Figure reprinted with permission from Yu et al., 2021 . Figure 5 Analysis of CD8 + T cell infiltration by immunofluorescence (A) Frozen sections of tumor tissue were stained for CD8 (green) and DAPI (blue).…”
Section: Expected Outcomesmentioning
confidence: 99%
“…Data are mean ± SEM, two tailed T test. Figure reprinted with permission from Yu et al., 2021 . Figure 6 Lentivirus-sucrose cocktail in ultracentrifuge tube Related to step 2b.…”
Section: Expected Outcomesmentioning
confidence: 99%
“… Representative images of flow cytometry analysis of CD8 + , GZMB + , and IFN-γ + Cells in TDCL tumors, CD45 + cells were isolated from subcutaneous primary tumor tissues (A), plots showing the mean percentage ± SEM of CD8 + T cells in CD45 + cells, GZMB + cells, and IFN-γ + cells in tumor treated with IgG and aPD-1(n=6 for each group), two-tailed t test (B). Figure reprinted with permission from Yu et al., 2021 . …”
Section: Expected Outcomesmentioning
confidence: 99%
“…This protocol can also be applied to other cancer types and immunotherapies. For complete details on the use and execution of this protocol, please refer to Yu et al. (2021) …”
Summary
Anti-PD-1/PD-L1 therapy shows long-term effects in many cancer types, but resistance and relapse remain the main limitations of this therapy. Here, we describe a protocol to evaluate the tumor response to immunotherapy in a mouse lung cancer model. The protocol includes the establishment of the lung cancer mouse model, anti-PD-1 treatment, tumor-infiltrating lymphocyte isolation, immunofluorescence, and flow cytometry analysis. This protocol can also be applied to other cancer types and immunotherapies.
For complete details on the use and execution of this protocol, please refer to
Yu et al. (2021)
Phase separation is a hot research field at present. It involves almost all aspects of cells and plays a significant role in cells, promising to be “a master key” in unlocking the mysteries of nature. In this review, the factors that affect phase separation are introduced, such as own component, electrostatic interaction, and chemical modification. Furthermore, the physiological roles of phase separation in cells, including molecules transport channel, gene expression and regulation, cellular division and differentiation, stress response, proteins refolding and degradation, cell connections, construction of skin barrier, and cell signals transmission, are highlighted. However, the disorder of phase separation leads to pathological condensates, which are associated with neurodegenerative diseases, tumors, and severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). This relationship is considered the potential target for developing corresponded drugs and therapy. Some drugs targeting phase separation have improved meaningful, such as tankyrase, lipoamide, oligonucleotides, elvitagravir, nilotinib, CVL218, PJ34. All in all, mystery phase separation provides a new viewpoint for researchers to explore cells, and is expected to solve many unknown phenomena in nature.
Induced pluripotent stem cells (iPSCs) have wide applications in disease modeling, personalized medicine, and tissue engineering. The generation of iPSCs from somatic cells via transcriptional‐factor‐ or chemical molecule‐based approaches are time‐consuming and inefficient. Here, a cell‐reprogramming‐inspired dynamically responsive hydrogel is fabricated via a synthetic‐biology‐based strategy. Human and mouse somatic cells (including senescent cells) are efficiently reprogrammed into iPSCs that exhibit key features of embryonic stem cells. The cell‐reprogramming‐responsive hydrogel possesses dynamic bioresponsiveness, and it faithfully senses metabolic remodeling and extracellular acidification during cell reprogramming, responding by changing its mechanical properties accordingly. Mechanistic study demonstrates that the autonomous change of the mechanical properties of the cell‐reprogramming‐responsive hydrogel elicits the formation of Yes‐associated protein (YAP) biomolecular condensates with the appropriate timing during cell reprogramming, ensuring a faster and more efficient generation of iPSCs than conventional cell reprogramming approach. Taken together, this study reveals the robust induction of pluripotency by coordination of cell‐reprogramming‐inspired dynamically responsive hydrogel and phase‐separated biomolecular condensates.
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