Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: Primer sequences for qPCR

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Sequencing

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: Knockdown of TPM3 inhibits the proliferation and induces the apoptosis of ESCC cell lines. (A) The TPM3 protein expression level was detected by western blot in shTPM3 ESCC cells. (B,C) CCK8 assays detected the proliferation of ESCC cells (n=3). (D) Colony formation assay analyzed the proliferation of ESCC cells, and the number of colonies stained with crystal violet was the basis for comparison (n=3). (E) The proportion of early apoptosis cells in ESCC cells was tested by FCM and Annexin V/PI double staining (n=3). *, ** represent P<0.05, P<0.01 respectively. ESCC, esophageal squamous carcinoma; CCK8, Cell Counting Kit-8; FCM, flow cytometry; PI, propidium iodide.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Knockdown, Expressing, Western Blot, Colony Assay, Staining, Comparison, Double Staining, Cell Counting, Flow Cytometry

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: TPM3 down-regulation weakens the invasion and migration of ESCC cell lines. (A) Transwell assays were used to evaluate the invasion and the migration of ESCC cells in vitro . The number of cells penetrating the filter membrane coated with matrigel revealed the invasion ability of cells, while the transwell without matrigel could detect the migration of cells (n=3). (B) In the wound-healing assay, the migration of cells was also analyzed, and the distance that cells moved was measured and normalized using the blank group (n=3). All cells were stained with crystal violet. *, ** represent P<0.05, P<0.01, respectively. ESCC, esophageal squamous carcinoma.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Migration, In Vitro, Membrane, Wound Healing Assay, Staining

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: TPM3 promotes ESCC cell proliferation and lung metastasis in vivo . (A) The growth curve of subcutaneous tumors in nude mice (day 30, shNC 505.92±72.49 mm 3 ; shTPM3 235.65±38.21 mm 3 ; n=5). (B) Comparison of the size and weight of the removed tumors (day 30, size of tumors: shNC 10.36±0.52 mm, shTPM3 9.14±0.45 mm; weight of tumors: shNC 0.71±0.14 g; shTPM3 0.33±0.07 g; n=5). (C) Detection results of Ki-67 and TUNEL in subcutaneous tumors, and IHC staining were used to detect Ki-67. (D) The tumor percentage in the lungs of the tail-vein injection nude mice metastasis model (day 30, shNC 80.41%±3.95%, shTPM3 37.80%±7.86%; n=5), and the paraffin sections were stained with HE. **, *** represent P<0.01, P<0.001, respectively. ESCC, esophageal squamous carcinoma; TUNEL, terminal deoxynucleotidyl transferase nick-end-labeling; IHC, immunohistochemistry; HE, hematoxylin/eosin.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: In Vivo, Comparison, TUNEL Assay, Immunohistochemistry, Injection, Staining, End Labeling

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: TPM3 and miR-107 expression in tissues and cell lines. (A, B) The qPCR detection results of TPM3 and miR-107 in human ESCC tissues and the paired adjacent normal tissues (n=15). (C, D) The qPCR detection results of TPM3 and miR-107 in human ESCC cell lines and HEEC. (E) The western blot image and the grayscale quantization result. *, **, and *** represent P<0.05, P<0.01, and P<0.001, respectively. qPCR, quantitative polymerase chain reaction; ESCC, esophageal squamous carcinoma; HEEC, human esophageal epithelial cells.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: MiR-107 inhibits TPM3 expression by targeting its 3’-UTR. (A) MiR-107 mimics and inhibitor could influence the expression of TPM3, which was confirmed by qPCR and western blot. (B) We designed the mutant sequence (TPM3-MUT) based on the predicted sequence (TPM3-WT) and compared the fluorescence intensity of miR-107 mimics and inhibitor intervention before and after the predicted binding region site mutation in the dual-luciferase reporter gene assay. *, **, and *** represent P<0.05, P<0.01, and P<0.001, respectively. 3’-UTR, 3’-untranslated region; qPCR, quantitative polymerase chain reaction.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Expressing, Western Blot, Mutagenesis, Sequencing, Fluorescence, Binding Assay, Luciferase, Reporter Gene Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: MiR-107 plays an anti-cancer role in ESCC and this effect could be rescued by TPM3. (A,B) Proliferation analysis of ESCC cells based on the CCK8 assay when the miR-107 mimics and exogenous plasmid containing TPM3 were applied (n=3). (C) The detection of early apoptosis levels based on the FCM assay in the case of miR-107 mimics and exogenous TPM3 plasmid intervention (n=3). (D) Transwell experiments results before and after the application of miR-107 mimics and TPM3 plasmid (n=3), here cells were stained with crystal violet. *, **, and *** represent P<0.05, P<0.01, and P<0.001, respectively. ESCC, esophageal squamous carcinoma; CCK8, Cell Counting Kit-8; FCM, flow cytometry.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: CCK-8 Assay, Plasmid Preparation, Staining, Cell Counting, Flow Cytometry

Journal: Journal of Gastrointestinal Oncology

Article Title: MiR-107 inhibits the malignant biological behavior of esophageal squamous cell carcinoma by targeting TPM3

doi: 10.21037/jgo-22-575

Figure Lengend Snippet: Diagram of miR-107 inhibition of esophageal squamous carcinoma progression by targeting TPM3 expression.

Article Snippet: The following antibodies were used: TPM3 (Proteintech, Wuhan, China), GAPDH (Abcam, Shanghai, China), and secondary antibodies (Bioworld, Nanjing, China).

Techniques: Inhibition, Expressing

Journal: Cell Reports Medicine

Article Title: Targeting severe acidity for tumor-activatable Interleukin-2 therapy

doi: 10.1016/j.xcrm.2025.102572

Figure Lengend Snippet: pH-responsive blocking and restoration of IL-2 receptor binding by UPS 5.3 (A) Schematic of the experimental design to assess the intrinsic interaction between UPS 5.3 micelles and proteins. (B) FPLC chromatograms showing individual proteins or UPS 5.3 /protein mixtures. (C) Molecular docking model of the DBA dimer (blue) with IL-2, highlighting predicted interaction sites. (D–F) Biolayer interferometry analysis of IL-2-Fc binding to IL-2Rα, IL-2Rβ, and IL-2Rγ, with and without UPS 5.3 NP encapsulation. (G) Schematic illustrating ex vivo assessment of IL-2-Fc binding to immune cells from various organs. (H) Quantification of IL-2-Fc binding to lymphocytes isolated from liver, lung, spleen, and tumor ( n = 5). Data are presented as mean ± SEM. See also and .

Article Snippet: InVivoMAb anti-mouse CD122 (IL-2Rβ) , BioXcell , Cat# BE0298 RRID: AB_2687820.

Techniques: Blocking Assay, Binding Assay, Encapsulation, Ex Vivo, Isolation

Journal: Immunity

Article Title: CD8 + T cell activation in cancer comprises an initial activation phase in lymph nodes followed by effector differentiation within the tumor

doi: 10.1016/j.immuni.2022.12.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: anti-human CD141 (FITC) , Miltenyi Biotec , clone: REA674, RRID: AB_2751167.

Techniques: Blocking Assay, Recombinant, Staining, Selection, Enzyme-linked Immunosorbent Assay, Software