Journal: Journal for immunotherapy of cancer

Article Title: Novel PAP-targeted CAR-T therapy enhances antitumor efficacy through CoupledCAR approach.

doi: 10.1136/jitc-2024-011238

Figure Lengend Snippet: Figure 5 The CoupledCAR approach enhanced the efficient expansion and enhanced the functionality of PAP CAR-T cells in vivo. (A) Schematic illustration of the in vivo studies. The prostate cancer xenograft model was established by the subcutaneous injection of 2.0×106 PC3-tmPAP cells, followed by the administration of different T and B cell or MOCK infusion strategies 24 days later (D0). The mice were divided into the MOCK group (n=7), NT group (n=7), B cell group (n=7), PAP CAR-T group (n=7), PAP CAR-T cell+CD19 CAR-T cell group (n=7), CD19 CAR-T cell group (n=7), CD19 CAR-T cell+B cell group (n=7), PAP CAR-T cell+B cell group (n=7) and PAP CAR-T cell+CD19 CAR-T cell+B cell group (n=7). A total of 1.0×106 B cells were infused on Day 3, 6 and 9 in the groups that received B-cell infusion. (B–G) Peripheral blood was collected on Day 14 to analyze CAR-T cell expansion (B, C) and cytokine release (D, E). The tumor volumes were measured every 3 days. (An unpaired t-test was used to compare between PAP CAR-T cell+CD19 CAR-T cell+B cell group and PAP CAR-T group, PAP CAR-T cell+CD19 CAR-T cell+B cell group and PAP CAR-T cell+CD19 CAR-T cell group, PAP CAR-T cell+CD19 CAR-T cell+B cell group and PAP CAR-T cell+B cell group on Day 15) (F). (G) The infiltration of CD3+, CD4+, CD8+, and PAP-CAR-transduced T cells into mouse tumors was assessed by immunohistochemistry. Scale bars represent 200 µm. (H–K). The histograms show the infiltration of CD3, CD4, CD8, and PAP CAR-T cells in tumor tissues. Intergroup comparisons in (B) were analyzed using an unpaired t-test. Intergroup comparisons in (C–F) and (H–K) were analyzed using one-way analysis of variance. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, and p≥0.05; NS, no significance. CAR, chimeric antigen receptor; GFP, green fluorescence protein; IFN, interferon; i.v., intravenous; NT, non-transduced; PAP, prostatic acid phosphatase; PB, peripheral blood; tmPAP, transmembrane PAP; s.c., subcutaneous.

Article Snippet: IHC analysis of tumor tissue sections obtained from animal experiments was performed using the following primary antibodies: CD3 (Cell Signaling Technology, #85061), CD4 (CST, #48274), CD8 (CST, #85336S), and GFP (CST, #2956S).

Techniques: In Vivo, Injection, Immunohistochemistry, Fluorescence

Journal: Biotechnology for Biofuels

Article Title: Improved cold tolerance in switchgrass by a novel CCCH-type zinc finger transcription factor gene, PvC3H72 , associated with ICE1–CBF–COR regulon and ABA-responsive genes

doi: 10.1186/s13068-019-1564-y

Figure Lengend Snippet: PvC3H72 subcellular localization and transcriptional activity assay. a Subcellular localization of PvC3H72–GFP fusion protein in Arabidopsis protoplast. GFP protein alone shows fluorescent signals in nucleus, membrane, and cytoplasm. DAPI is a staining dye for the nucleus. Bars = 5 µm. b PvC3H72 transcriptional activity in yeast system. GUS was used as a negative control. c Schematic diagram of three plasmids used for transactivation activity. Three plasmids (effector, reporter and internal control) were electroporated into Arabidopsis protoplasts at the ratio of 5:4:1. d PvC3H72 showed transactivation activity in the plant cells. All the experiments were repeated three times with similar results. Letters above bars indicate significant difference at P < 0.05 ( n = 3)

Article Snippet: DAPI were used to stain the nucleus, and the GFP signal were detected under a Zeiss LSM 780 laser scanning confocal microscope (Carl Zeiss SAS, Jena, Germany).

Techniques: Activity Assay, Membrane, Staining, Negative Control, Control