mouse α stat1  (Cell Signaling Technology Inc)

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: CD40 agonism triggers a metabolic reprogram of TAMs similar to that of Trem2 -KO (A) Heatmap of differential gene set analysis in TAM subpopulations. Genes were enriched in Mac_Cxcl9 cluster. (B) The network diagram illustrated the highly expressed genes in Mac_Cxcl9 cluster and their interactions with shared transcription factors. (C) Correlation analysis between the ratio of Cd40 + TAMs and Cxcl9 + TAMs . Each dot represented a single-cell sequencing sample of tumor and spleen. (D) BMDMs isolated from Trem2 -KO or WT mice were educated for 24 h in a transwell system with cancer cells, with or without IFN-γ intervention. Relative protein expression levels of CD40 were assessed by WB and normalized to β-actin. (E) Schematic representation of in vitro tumor-educated macrophage model. BMDMs isolated from WT mice were educated for 24 h in a transwell system with cancer cells, with or without IFN-γ (40 ng/mL) and CD40 agonism (crosslinked FGK45, 20 ng/mL) intervention. (F) Relative mRNA expression levels of Cxcl9 and Spp1 in BMDMs were quantified by RT-qPCR, and the ratio of Cxcl9/Spp1 was calculated. n = 3. (G and H) The real-time changes of OCR of BMDMs were stimulated with or without IFN-γ and CD40 agonism in the basal state and following the additions of oligomycin (Oligo), FCCP, etomoxir (Eto), and rotenone + antimycin A (Rot/AA). The average of basal OCR, maximal OCR, and Eto-sensitive OCR were revealed. n = 3. (I and J) The real-time measurement of ECAR of BMDMs were stimulated with or without IFN-γ and CD40 agonism in the basal state and following the additions of glucose (Gluc), oligomycin (Oligo), and 2-deoxy-D-glucose (2-DG). The average of basal ECAR, maximal ECAR, and glycolytic reserve were revealed. n = 3. (K, L, O, and P) BMDMs from WT mice were co-cultured with tumor cells in transwells for 24 h with or without IFN-γ (40 ng/mL) and CD40 agonism (crosslinked FGK45, 20 ng/mL) (K and O). For inhibitor studies, BMDMs were pre-treated with AMPK inhibitor (20 μM) (L) or STAT1 inhibitor (100 μM) (P) for 2 h prior to co-culture. Protein levels were analyzed by western blot and normalized to β-actin. (M and N) AMP and ATP concentrations in BMDMs were measured using dedicated assay kits, and the AMP/ATP ratio was calculated from these values. n = 3. (Q) Mechanism schema diagram. Data were represented by mean ± SEM. ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: STAT1 α Rabbit mAb , CST , Cat. #9172.

Techniques: Sequencing, Isolation, Expressing, In Vitro, Quantitative RT-PCR, Cell Culture, Co-Culture Assay, Western Blot

Journal: Infection and Immunity

Article Title: TRP75-mediated STAT3 activation promotes anti-apoptotic signaling and Ehrlichia chaffeensis infection

doi: 10.1128/iai.00459-25

Figure Lengend Snippet: Nuclear translocation of STAT family members during E. chaffeensis infection. ( A ) Uninfected or E. chaffeensis -infected THP-1 cells at 72 hpi (MOI 10) were probed for STAT1–6 (green) and E. chaffeensis marker Dsb (red) by immunofluorescent microscopy. 4´,6-Diamidino-2-phenylindole (DAPI) (blue) was used to stain nuclei. Scale bar = 20 µm. ( B ) Nuclear regions of interest (ROIs) were defined by DAPI signal, and mean nuclear STAT3 intensity per cell was quantified using ImageJ. Statistical significance was determined using two-tailed Student’s t -test; * P < 0.05, ** P < 0.01. Data represent the mean ± SD of three independent biological replicates ( n = 3), and representative images are shown.

Article Snippet: The primary antibodies utilized in this research include monoclonal rabbit α-STAT1 (14994S; Cell Signaling Technology), monoclonal rabbit α-STAT2 (72604S; Cell Signaling Technology), rabbit α-STAT3 (12640S; Cell Signaling Technology), monoclonal rabbit α-STAT4 (2653S; Cell Signaling Technology), monoclonal rabbit α-STAT5 (94205S; Cell Signaling Technology), monoclonal rabbit α-STAT6 (5397S; Cell Signaling Technology), monoclonal rabbit α-phospho-STAT3 Y705 (9145S; Cell Signaling Technology), monoclonal rabbit α-MCL-1 (94296S; Cell Signaling Technology), monoclonal rabbit α-caspase 3 (9662S; Cell Signaling Technology), monoclonal mouse α-caspase 7 (9494S; Cell Signaling Technology), monoclonal mouse α-GAPDH (sc-47724; Santa Cruz Biotechnology), monoclonal mouse α-Tubulin (sc-5286; Santa Cruz Biotechnology), monoclonal mouse α-Vinculin (sc-73614; Santa Cruz Biotechnology), monoclonal mouse α-GFP (sc-9996; Santa Cruz Biotechnology), rabbit α-disulfide bond formation protein (Dsb) , and rabbit α-TRP75 ( ).

Techniques: Translocation Assay, Infection, Marker, Microscopy, Staining, Two Tailed Test

Journal: Scientific Reports

Article Title: Polystyrene microplastics are internalized by human gingival fibroblasts, enhance cell motility and induce molecular changes revealed through proteomic analysis

doi: 10.1038/s41598-025-19064-w

Figure Lengend Snippet: Validation of proteomic data by Western blotting. Representative Western blot analysis showing the expression of Fibronectin ( A ), STAT1 ( B ), CALR ( C ), and VDAC1 ( D ) in hGF cells after 48 and 72 h of PS-MPs treatment. Cells cultured in medium alone served as negative control (CTRL). β-actin was used as a loading reference protein. Quantification of the relative protein expression of Fibronectin ( A ), STAT1 ( B ), CALR ( C ), and VDAC1 ( D ) in hGF cells after 48 and 72 h of PS-MPs treatment, normalized to CTRL. (* p -value < 0.05).

Article Snippet: Proteins were transferred onto nitrocellulose membrane, blocked for 1 h with 5% BSA and 1% non-fat dried milk in Tris-Buffered Saline (TBS), and incubated overnight at 4 °C with primary antibodies: α-fibronectin (#F3648, Sigma-Aldrich, MO, USA), α-STAT1 (#9172, Cell Signaling, MA, U.S.A), α-Calreticulin (#SPA-601, Enzo Life Sciences, Inc., NY, U.S.A), α-VDAC1 (#PA1954A, Invitrogen, MA, USA), and α-β-actin (#sc-69879, Santa Cruz, CA, USA).

Techniques: Biomarker Discovery, Western Blot, Expressing, Cell Culture, Negative Control