Journal: International Journal of Nanomedicine

Article Title: Photothermal exposure of polydopamine-coated branched Au–Ag nanoparticles induces cell cycle arrest, apoptosis, and autophagy in human bladder cancer cells

doi: 10.2147/IJN.S174349

Figure Lengend Snippet: Changes of ΔΨm and the release of cytochrome c in T24 cells treated with different concentrations of Au–Ag@PDA NPs at 3 hours after laser irradiation with 1 W/cm 2 for 4 minutes. Notes: ( A ) The ΔΨm of T24 cells was determined by FCM using JC-1 staining. ( B ) Red/green fluorescence intensity ratios. ( C ) Level of cytosolic cytochrome c measured by Western blot. GAPDH was used as the loading control. (0-: 0 µg+no laser, 0: 0 µg+laser, 20: 20 µg+laser, 30: 30 µg+laser, 40: 40 µg+laser). ( D ) Quantification analysis of cytochrome c /GAPDH compared with the 0- control group (mean±SD; n=3). * P <0.05 vs 0 group. Abbreviations: ΔΨm, mitochondrial membrane potential; FCM, flow cytometry; NPs, nanoparticles; PDA, polydopamine.

Article Snippet: In brief, the cells were trypsinized, incubated with JC-1 solution at 37°C for 20 minutes, washed twice with PBS, and then analyzed using FCM (FACSCanto II; Becton, Dickinson and Company, Franklin Lakes, NJ, USA).

Techniques: Irradiation, Staining, Fluorescence, Western Blot, Flow Cytometry

Journal: Cellular and Molecular Immunology

Article Title: CD2 augmentation enhances CAR-T-cell efficacy via immunological synapse remodeling and T-cell exhaustion mitigation

doi: 10.1038/s41423-025-01314-6

Figure Lengend Snippet: CD2 is decisive for the quality of CAR-IS. A Immunofluorescence imaging (left) and colocalization analysis (right) of CD2 (green) and CAR (red) in Jurkat CD19-BBζ CAR-T cells. The data are presented as Pearson’s correlation coefficient (PCC) values ( n = 10; mean ± SEM; scale bar = 5 μm). B Confocal microscopy of Jurkat CAR-T cells in combination with Raji cells (left). Raji cells were labeled with CellTrace™ Violet dyes. Fixed and permeabilized cells were stained with F-actin (TRITC Phalloidin). The IS was evaluated by measuring the ratio of F-actin in and out of synapses (right) ( n = 30; mean ± SEM; scale bar = 10 μm; unpaired t test). C Confocal microscopy of Jurkat CAR-T cells (J-CD19-BBζ), Jurkat CD2 null CAR-T cells (J-CD2KO-CD19-BBζ), J-CD2KO-CD19-BBζ cells expressing the extracellular domain of CD2 (J-CD2KO-CD19-BBζ + CD2TL) and J-CD2KO-CD19-BBζ cells expressing full-length CD2 (J-CD2KO-CD19-BBζ + CD2) in conjugation with Raji cells. Raji cells were labeled with CellTrace™ Violet dyes. TRITC-phalloidin was used to stain F-actin. The IS was evaluated by measuring the ratio of F-actin in and out of synapses (right) ( n = 30; mean ± SEM; scale bar = 10 μm; ordinary one-way ANOVA). D CAR ligation initiates proximal signaling events in J-CD19-BBζ (marked as 19), J-CD2KO-CD19-BBζ (marked as KO), J-CD2KO-CD19-BBζ + CD2 (marked as CD2) and J-CD2KO-CD19-BBζ + CD2TL (marked as TL) cells. E Proximal signaling events in Jurkat CAR-T cells upon stimulation with Nalm6 WT or Nalm6 CD58KO cells. F Activation signals of J-CD19-BBζ (marked as 19) and J-CD19-BBζ + CD2 (marked as CD2) CAR-T cells cocultured with CD19 − CD58⁺ K562 cells. The image is uniformly brightness/contrast adjusted; see supplementary materials for raw data * p < 0.05, ** p < 0.01

Article Snippet: After three washes with PBS, the cells were incubated with 200 μL of prepared TRITC-labeled phalloidin working solution (Yeasen) in the dark at room temperature for 30 min.

Techniques: Immunofluorescence, Imaging, Confocal Microscopy, Labeling, Staining, Expressing, Conjugation Assay, Ligation, Activation Assay

Journal: Cellular and Molecular Immunology

Article Title: CD2 augmentation enhances CAR-T-cell efficacy via immunological synapse remodeling and T-cell exhaustion mitigation

doi: 10.1038/s41423-025-01314-6

Figure Lengend Snippet: CD2-CAR-T cells demonstrate enhanced antitumor potential. A The quality of the IS formed by J-CD19-BBζ and J-CD19-BBζ + CD2 CAR-T cells conjugated with Nalm6 cells ( n = 10; mean ± SEM; unpaired t test). B The percentages of J-CD19-BBζ and J-CD19-BBζ + CD2 CAR-T cells remaining bound to Nalm6 cells at 1000 pN ( n = 3; mean ± SEM; unpaired t test). C The proportion of live Raji cells during the 6-h coculture with CAR-T cells at an E : T ratio of 1:10 (left). The proportion of PKH67+ Raji cells that contacted CAR-T cells among live Raji cells (right). D The expression level of CD19 on live Raji cells ( n = 3; mean ± SEM; two-way ANOVA). E Lysis efficiency of Nalm6, Raji and A549-CD19 CAR-T cells treated with CD19-BBζ and CD19-BBζ + CD2 CAR-T cells at the indicated E : T ratios for 24 h ( n = 3; mean ± SEM; two-way ANOVA). F Concentrations of cytokines released by CD19-BBζ- and CD19-BBζ + CD2 CAR-T cells after 24 h of coculture with Raji cells at an E : T ratio of 1:10. ( n = 3; mean ± SEM; unpaired t test). G CD2 expression levels in CD19-BBζ and CD19-BBζ + CD2 CAR-T cells after each stimulation with Nalm6 cells at an E : T ratio of 1:1. ( n = 3; mean ± SEM). H Lysis efficiency of CD19-BBζ CAR-T cells and CD19-BBζ + CD2 CAR-T cells against Nalm6 cells at an E : T ratio of 1:1 for 24 h in each round of coculture ( n = 3; mean ± SEM; two-way ANOVA). I Lysis efficiency of CD19-BBζ and CD19-BBζ + CD2 CAR-T cells against Raji cells at an E : T ratio of 1:5 for 24 h in each round of coculture ( n = 3; mean ± SEM; two-way ANOVA). J Confocal microscopy of CD19-BBζ and CD19-BBζ + CD2 CAR-T cells (before and after three rounds of stimulation) stimulated with Raji cells. Raji cells were labeled with CellTrace™ Violet dyes. TRITC-phalloidin was used to stain F-actin. The IS was quantified by measuring the fluorescence density ratio of F-actin in and out of synapses (left) ( n = 11, mean ± SEM; scale bar = 10 μm). * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: After three washes with PBS, the cells were incubated with 200 μL of prepared TRITC-labeled phalloidin working solution (Yeasen) in the dark at room temperature for 30 min.

Techniques: Expressing, Lysis, Confocal Microscopy, Labeling, Staining, Fluorescence

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuroprotective Effects of Transcranial Pulsed Current Stimulation: Modulation of Microglial Polarization in Traumatic Brain Injury

doi: 10.1111/cns.70606

Figure Lengend Snippet: Protective effects of tPCS on neurological function, pathological alterations, and inflammatory response following TBI. (A) Experimental procedure timeline. (B) Neurological function in mice was assessed using the mNSS at 1, 3, 5, and 7 dpo ( n = 8). (C) Representative exploration routes in the Y maze. (D) Time spent in the novel arm of the Y‐maze ( n = 8). (E) Beam‐balance test measuring the time required to traverse the beam ( n = 8). (F) Rotarod test measuring the latency to fall ( n = 8). (G) Representative scanning images and photomicrographs of H&E‐stained sections at 7 dpo ( n = 4). Arrows point to the neuronal edema, vacuolation, and degeneration. (H) Representative scanning images and photomicrographs of Nissl‐stained sections at 7 dpo ( n = 4). Arrows highlight neuronal vacuolation and degeneration. (G) Quantification of Nissl‐stained cells. (I–K) Quantitative analysis of Nissl‐stained cells, lesion volume, and correlation analysis between lesion volume and mNSS. (L) Representative images of TUNEL staining at 7 dpo. (M) Quantification of TUNEL‐stained cells ( n = 4). (N–P) The levels of inflammatory mediators including IL‐1β, TNF‐α, and IL‐10 in the peri‐lesional cortex were detected by ELISA ( n = 4). Scale bar = 100 μm. All values are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After dehydration and dewaxing, sections were incubated with TUNEL working solution (Servicebio, China) at 37°C for 2 h, followed by DAPI staining at 4°C for 8 min. After thorough rinsing, the sections were dried, mounted, and examined under a fluorescence microscope.

Techniques: Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Atorvastatin promotes lipid catabolism in colorectal cancer via FDFT1‒mediated inhibition of the PI3K/AKT pathway

doi: 10.1186/s12964-026-02802-6

Figure Lengend Snippet: ATST induces ER stress and promotes apoptosis in CRC cells A Expression of ER stress‒related proteins in HCT116 cells after ATST treatment B Changes in intracellular Ca 2+ levels in HCT116 cells C Alterations in XBP‒1s mRNA expression in HCT116 cells D Representative IF images of XBP‒1s in HCT116 cells after ATST treatment. Scale bar: 50 μm E Apoptosis rate in HCT116 cells after ATST treatment F Changes in JC‒1 ratio in HCT116 cells following ATST treatment G Expression of apoptosis‒related proteins in HCT116 cells after ATST treatment H Representative IF images of CHOP in HCT116 cells after ATST treatment. Scale bar: 50 μm I Expression of SQSTM1 and LC3 A/B proteins in HCT116 cells after ATST treatment J Change in ROS levels in HCT116 cells K Cell cycle distribution in HCT116 cells after ATST treatment

Article Snippet: Cells were seeded in 6‒well plates at a density of 5 × 105 cells per well and treated with ATST at specified concentrations for 48 h. Cells were then collected and incubated with JC‒1 working solution (Yesen, 717164ES03, Shanghai) at 37 °C for 20 min. After washing twice with serum‒containing medium, samples were analysed by flow cytometry.

Techniques: Expressing