Journal: Molecular Medicine

Article Title: The SARS-CoV-2 nucleocapsid protein induces microglia senescence-mediated cognitive impairment via Glycolysis

doi: 10.1186/s10020-025-01410-3

Figure Lengend Snippet: SARS-CoV-2 N protein drives the senescence of BV2 microglial cells by triggering mitochondrial dysfunction. BV2 microglial cells were treated with Mdivi-1 (100 nM) 30 min before Codon-optimized pLJM1-SARS-CoV-2 N-FLAG transfection. A Representative image of BV2 cells loaded with the mitochondrial membrane potential indicator JC-1 (bar = 20 μm). B Mitochondrial morphology stained with TOM20 in BV2 cells (bar = 2 μm). C - D The expression levels of p-DRP1 and DRP1 proteins in BV2 microglial cells detected by Western blot ( n = 3). Bar graph data represent the mean ± SD of the target protein intensity (normalized to DRP1) in the experimental group relative to that in the control group. E Representative images of BV2 cells loaded with the mitochondrial membrane potential indicator JC-1 (bar = 20 μm). F The expression levels of p19 , p21 , p53 , Il-1β , and Tnf-α mRNA in BV2 microglial cells detected by real-time PCR ( n = 3). G SA-β-gal staining was performed after transfection of Codon-optimized pLJM1-SARS-CoV-2 N-FLAG and treatment with Mdivi-1 for 36 h (bar = 10 μm). H The fluorescence intensity of Ki67 (green) was detected by immunofluorescence (bar = 50 μm). I - J The expression levels of p-DRP1, p53, p21, p16, and γ-H2AX proteins in BV2 microglial cells were detected by Western blot ( n = 3). Bar graph data represent the mean ± SD of the target protein intensity (normalized to α-tubulin) in the experimental group relative to that in the control group. Comparisons between the two groups were made with an unpaired t -test. Differences among multiple groups were performed using ANOVA. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Rabbit-anti-p-DRP1 S616 polyclonal antibody , CST , 3455 , 1:2000.

Techniques: Transfection, Membrane, Staining, Expressing, Western Blot, Control, Real-time Polymerase Chain Reaction, Fluorescence, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: TGF-β1 suppressed OXPHOS activity in CD4 + human T cells (A–D) CD4 + and CD8 + T cells (1 × 10 6 ) were activated with CD3/CD28 mAbs for 24 h, followed by PBS or TGF-β1 (10 ng/mL) treatment for 16 h. (A) Mitochondrial morphology of CD4 + and CD8 + T cells upon PBS or TGF-β1 (10 ng/mL) treatment, as determined by spinning disk confocal microscopy. Mitochondria are green (MitoTracker Green), and nuclei are blue (DAPI). Scale bar, 5 μm. (B) Relative lengths of mitochondria, as analyzed by ImageJ software, in CD4 + and CD8 + T cells (2 independent experiments). Each dot represents the mean relative length of the mitochondria in a sample. Data are shown as the mean ± SEM values; paired two-tailed t test; ∗∗∗∗ p ≤ 0.0001. (C) Immunoblot analysis of cellular protein extracts probed with antibodies against pSMAD2 S465/467 (top)/pSMAD3 S423/425 (bottom) (pSMAD2/3), SMAD2/3, OPA1, MFF, pDRP1 S616 , DRP1, and β-actin. (D) The relative expression of pDRP1 S616 , MFF, and OPA1 was analyzed by ImageJ software (3 independent experiments). Data are shown as the mean ± SEM values; paired two-tailed t test; ∗ p < 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. (E) Immunoprecipitation (IP) of MFF in activated CD4 + T cells after treatment with PBS or TGF-β1 (10 ng/mL) for 2 h and subsequent immunoblot (IB) analysis of the indicated proteins. (F) Mitochondrial morphology of CAR + CD4 + T28zT2, CAR + CD4 + G28zT2, and untransduced CD4 + T cells upon treatment with PBS or TGF-β1 (10 ng/mL) as determined by spinning disk confocal microcopy. Mitochondria are red (MitoTracker Deep Red), CAR T cells are green (GFP), and nuclei are blue (Hoechst 33342). Scale bar, 5 μm. (G) Lengths of mitochondria, as analyzed by ImageJ software (2 independent experiment), in CAR + CD4 + T28zT2, CAR + CD4 + G28zT2, and untransduced CD4 + T cells. Each dot represents the mean relative length of the mitochondria in a sample. (H–M) OCR profile (H and K), ATP-coupled OCR (I and L), and SRC (J and M) of CAR + CD4 + T28zT2 T cells and CAR + CD4 + G28zT2 T cells (3 independent experiments). Data are shown as the mean ± SEM values; unpaired two-tailed t test; ∗ p < 0.05, ∗∗ p ≤ 0.01. See also in Figure S6 .

Article Snippet: SMAD2/3 (clone D7G7) rabbit mAb, phospho-SMAD2 (S465/467)/SMAD3 (S423/425) (clone D27F4) (pSMAD2/3) rabbit mAb, SMAD4 (clone D3R4N) rabbit mAb, DRP1 (clone D6C7) rabbit mAb, phospho-DRP1(Ser616) (pDRP1 S616 ) rabbit mAb (clone D9A1), phospho-DRP1 S616 rabbit polyclonal antibody (#3455), OPA1 (D7C1A) rabbit mAb and MFF (E5W4M) XP rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques: Activity Assay, Confocal Microscopy, Software, Two Tailed Test, Western Blot, Expressing, Immunoprecipitation

Journal: Cell Reports Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet:

Article Snippet: SMAD2/3 (clone D7G7) rabbit mAb, phospho-SMAD2 (S465/467)/SMAD3 (S423/425) (clone D27F4) (pSMAD2/3) rabbit mAb, SMAD4 (clone D3R4N) rabbit mAb, DRP1 (clone D6C7) rabbit mAb, phospho-DRP1(Ser616) (pDRP1 S616 ) rabbit mAb (clone D9A1), phospho-DRP1 S616 rabbit polyclonal antibody (#3455), OPA1 (D7C1A) rabbit mAb and MFF (E5W4M) XP rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques: Purification, Functional Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Cell Isolation, Western Blot, Software

Journal: Cell Communication and Signaling : CCS

Article Title: Impact of POU3F4 mutation on cochlear development and auditory function

doi: 10.1186/s12964-025-02133-y

Figure Lengend Snippet: Antibody reagents list for experimental studies

Article Snippet: Rabbit polyclonal anti-DRP1 , Proteintech , Cat#12957-1-AP.

Techniques: