Journal: Pharmaceutics

Article Title: Overcoming Resistance to Immune Checkpoint Inhibitor Therapy Using Calreticulin-Inducing Nanoparticle

doi: 10.3390/pharmaceutics15061693

Figure Lengend Snippet: CRT-NPs showed efficient plasmid encapsulation, stability, and transfection efficiency. ( A ) CRT-NPs showed an increased hydrodynamic diameter and a reduced zeta potential compared to blank liposomes, with excellent stability in the physiological buffer for 3 days. ( B ) TEM images (scale bar—500 nm) of CRT-NPs stored in physiological buffer and obtained over four consecutive days demonstrated excellent morphological stability. ( C ). Agarose gel imaging showed that the CRT plasmid (pCRT) remained bound to the NPs, as no free pCRT was observed in the unlysed CRT-NP lane. In contrast, the released plasmid from triton-lysed CRT-NPs (~2.6 µg) exhibited a band corresponding to a 2.5 µg free pCRT band. ( D ) The transfection efficiency of CRT-NPs was confirmed using EGFP as a model plasmid. Significant GFP expression (green) was observed with transfected EGFP-NP CT26 cells at 48 h compared to the untreated control, unencapsulated NP, and free EGFP plasmid groups. ( E ) CRT-NPs induced cell death in transfected CT26 cells in a dose-dependent manner. ( F ) Transfected CRT-NP cells showed significant increase in CRT gene expression when compared to other groups in qRT-PCR. ( G ) CRT-NP-induced apoptosis was significant different compared to untreated control and comparable to positive controls such as DOX and LF™2000 + pCRT-treated cells. The statistical significance was indicated as * p < 0.05, ** p < 0.005, **** p < 0.0001.

Article Snippet: To visualize the transfection efficacy of NPs, CT26 cells were transfected with EGFP-NP lipoplexes (CLs/pDNA) ( D).

Techniques: Plasmid Preparation, Encapsulation, Transfection, Zeta Potential Analyzer, Liposomes, Agarose Gel Electrophoresis, Imaging, Expressing, Control, Gene Expression, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Regulated Mesenchymal Stem Cells Mediated Colon Cancer Therapy Assessed by Reporter Gene Based Optical Imaging

doi: 10.3390/ijms19041002

Figure Lengend Snippet: Bystander effect of MSC-Tet-TK and MSC-TK cells. ( A ) Rluc activity in co-cultures (1:1) of naive MSCs and CT26/Rluc cells treated with the indicated concentrations of GCV for 48 h. ( B ) BLI images of the Rluc activity and quantitation data of CT26/Rluc in co-cultures (1:1) of MSC-TK or MSC-Tet-TK cells in the absence or presence of doxycycline (DOX(−) and DOX 2 μg/mL respectively). Three individual experiment values are expressed as the mean ± standard deviation (SD), * p < 0.05, ** p < 0.01, *** p < 0.001 (by Student’s t test). p/s, photons/second.

Article Snippet: CT26 cells were transduced with lentiviral particles expressing mCherry-Rluc (Renilla luciferase) under the control of the cytomegalovirus (CMV) promoter (Genecopoeia, Rockville, MD, USA).

Techniques: Activity Assay, Quantitation Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Regulated Mesenchymal Stem Cells Mediated Colon Cancer Therapy Assessed by Reporter Gene Based Optical Imaging

doi: 10.3390/ijms19041002

Figure Lengend Snippet: In vivo therapeutic effect of MSC-Tet-TK and MSC-TK cells on inhibiting colon tumor growth. ( A ) Renilla luciferase (Rluc) imaging of colon cancer cells (CT26/Rluc) in mice treated with either MSC-TK or MSC-Tet-TK cells with or without concurrent GCV treatment. BLI images were taken on days 0, 6, and 13 in five individual mice; ( B ) Quantitative analysis of the data shown in ( A ). ( C ) Tumor weights assessed at study end. Bioluminescence activity is shown in photons/second (p/s). * p < 0.05 compared separately to MSC-Tet-TK (GCV−) and MSC-TK (GCV−).

Article Snippet: CT26 cells were transduced with lentiviral particles expressing mCherry-Rluc (Renilla luciferase) under the control of the cytomegalovirus (CMV) promoter (Genecopoeia, Rockville, MD, USA).

Techniques: In Vivo, Luciferase, Imaging, Activity Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: XBP1 regulates the protumoral function of tumor-associated macrophages in human colorectal cancer

doi: 10.1038/s41392-021-00761-7

Figure Lengend Snippet: Effect of XBP1 activation on the pro-tumor function of TAMs. a Western blot analysis of XBP1s expression in indicated macrophages. b Representative micrograph showing tumor formation in NOD/SCID mice injected subcutaneously (s.c.) with luciferase tumor cells (CT26-luciferase) and two groups of macrophages: CT26 + BMDMs (black arrows) and CT26 + miTAMs (magenta arrows). c Growth curves of the two groups in b . d Western blot analysis of XBP1 expression in sgCon or sgXbp1 TAMs. e Representative photograph showing tumor formation in NOD/SCID mice injected s.c. with luciferase tumor cells (CT26-luciferase) and two groups of miTAMs: CT26 + sgCon miTAMs (black arrows); and CT26 + sgXbp1 miTAMs (blue arrows). f Growth curves of the two groups in e . **** P < 0.0001; Repeated measurement and analysis. g CT26 cells, mixed with: Con TAMs; Xbp1s TAMs; sgCon TAMs; and sgXbp1 TAMs, were orthotopically injected into the wall of the cecum ( n = 5 mice per group). Macroscopic appearance of the CRC orthotopic tumors with each indicated treatment. Black arrows indicate macroscopic polyps. Scale bar, 1 cm. h Representative HE staining of liver metastasis in mice xenografted of the four groups in g . Scale bar, 100 μm. i , j Statistical analysis of the orthotopic CRC tumor weight ( i ) and the clone number of liver metastasis ( j ). * P < 0.05, *** P < 0.001; t -test. k Incidence of orthotopic CRC tumor formation and liver metastasis analysis. l Representative images of CT26 pulmonary metastases induced by tail vein injection in NOD/SCID. CT26 cells were mixed with: Con miTAMs; Xbp1s miTAMs; sgCon miTAMs; and sgXbp1 miTAMs. HE staining demonstrating the histology of tumors formed in the lungs; scale bar, 500 μm. m Pulmonary metastatic nodule numbers in i . ( n = 4 mice per group); ** P < 0.01, *** P < 0.001; t -test

Article Snippet: Ana-1 (EP-CL-0023, RRID:CVCL_0142) and CT26 cells (EP-CL-0071, RRID:CVCL_7254) were from Elabscience (Texas, USA).

Techniques: Activation Assay, Western Blot, Expressing, Injection, Luciferase, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: XBP1 regulates the protumoral function of tumor-associated macrophages in human colorectal cancer

doi: 10.1038/s41392-021-00761-7

Figure Lengend Snippet: Effect of XBP1 on macrophages phagocytosis. a Representative images of phagocytosis assays using RFP-labeled human CRC cell, DLD1cells (DLD1-RFP) and sgCon or sgXBP1 hiTAMs ( n = 3). Yellow arrows denote phagocytic events. Scale bar = 200 μm. b Representative images of phagocytosis assays using RFP-labeled mouse CT26 cells (CT26-RFP) and sgCon or sgXbp1 miTAMs ( n = 3). Yellow arrows denote phagocytic events. Scale bar = 200 μm. c Results of phagocytosis assays of the two groups in a , b . ** P < 0.01; t -test. d Relative mRNA levels of XBP1 and phagocytosis-associated genes in BMDM, sgCon miTAMs and sgXbp1 miTAMs, validated by RT-qPCR. Bars represent mean ± SD of three experimental replicates. * P < 0.05, ** P < 0.01, *** P < 0.001; t -test. e Expression of THBS1 and SIRPα versus XBP1 in TAMs sorted from CRC patients ( n = 27 total). r , Spearman’s rank correlation test. f Correlation of XBP1 with THBS1 and SIRPα in CRC patients. The association was analyzed using coefficient measures of the linear relationships in the public GEO database (GSE68468). g Track view of Erdj4 , Thbs1 , and Sipra ChIP-seq density upon silencing of Xbp1 in the ChIP-seq online database (GSE86048). h ChIP-qPCR experiments measuring XBP1 binding on Erdj4 , Thbs1 , and Sipra segments. Bars represent mean ± SD of three experimental replicates. * P < 0.05, ** P < 0.01. P -values were determined using t -test

Article Snippet: Ana-1 (EP-CL-0023, RRID:CVCL_0142) and CT26 cells (EP-CL-0071, RRID:CVCL_7254) were from Elabscience (Texas, USA).

Techniques: Labeling, Quantitative RT-PCR, Expressing, ChIP-sequencing, ChIP-qPCR, Binding Assay

Journal: Frontiers in Veterinary Science

Article Title: Detection of bimodal survivin expressions in canine cancer types by flow cytometry compared to immunohistochemistry

doi: 10.3389/fvets.2025.1552415

Figure Lengend Snippet: Analysis of survivin expression sites by IHC and WB. (A) Images of HE staining and survivin expression via IHC in canine melanoma cell lines (CMM2, CMeC, and LMeC) are shown. (B) Images of HE staining and survivin expression via IHC in murine cell lines (p815, CT26, and B16F10) are shown. (C) The cytosol and total survivin expression patterns via WB are indicated. (D) The expression levels of total survivin (blue bar) and cytosolic survivin (orange bar) corrected on the basis of β-actin expression are shown via density of plot analysis via ImageJ software.

Article Snippet: A total of six cell lines were used: canine malignant melanoma lines [CMM2, CMeC2, LMeC; provided by Dr. Takayuki Nakagawa, Department of Veterinary Surgery, University of Tokyo; ( )], the murine malignant melanoma line B16F10, the murine mast cell tumor line p815, and the murine colon cancer line CT26 (distributed for a fee by the JCRB Cell Bank).

Techniques: Expressing, Staining, Software