Journal: Journal of Translational Medicine

Article Title: FCRLB-mediated dual control of tumor metabolism and macrophage polarization promotes lung cancer malignancy

doi: 10.1186/s12967-026-07872-1

Figure Lengend Snippet: Effects of FCRLB knockdown on ROS levels, CCL2 secretion, PI3K-AKT pathway activation, and macrophage polarization in non-small cell lung cancer (NSCLC) cells. ( A ) Reactive oxygen species (ROS) levels in two NSCLC cell lines transduced with control shRNA (shCtrl), shFCRLB-1#, or shFCRLB-2# were detected using a fluorescence microplate reader at an excitation wavelength (Ex) of 510 nm and an emission wavelength (Em) of 610 nm ( n = 3 per group). ( B ) ELISA was performed to measure the expression level of CCL2 in the cell supernatant of two NSCLC cell lines transduced with shCtrl, shFCRLB-1#, or shFCRLB-2# ( n = 6 per group). ( C – D ) WB analysis was used to detect the protein levels of key components in the PI3K-AKT signaling pathway in two NSCLC cell lines transduced with shCtrl, shFCRLB-1#, or shFCRLB-2#. Corresponding quantitative analyses of the p-PI3K/PI3K and p-AKT/AKT ratios are presented ( n = 3 per group). ( E ) Flow cytometry was employed to assess the effect of cell supernatant from LLC cells transduced with shCtrl or shFCRLB on the polarization of primary macrophages ( n = 3 per group). All error bars represent standard deviation (SD). *** P ≤ 0.001, ** P ≤ 0.01, * P ≤ 0.05

Article Snippet: After blocking, the following primary antibodies: FCRLB (Proteintech, 17157-1-AP), β-actin (Cell Signaling, 4967 S), p-PI3K (Cell Signaling,17366), p-AKT (Cell Signaling, 13038 S), mTOR (Cell Signaling, 2972 S) were used to incubate the membrane overnight.

Techniques: Knockdown, Activation Assay, Transduction, Control, shRNA, Fluorescence, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Standard Deviation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Multiomics Profiling Reveals Distinct Immunosuppression and Metabolic Dysregulation in Aggressive Subtypes of Thyroid Cancer

doi: 10.1016/j.mcpro.2026.101513

Figure Lengend Snippet: Identification of potential biomarkers in ATC/PDTC. A , volcano plot of DEPs in ATC/PDTC versus PTC with |log 2 FC| ≥ 1 and p < 0.05. B , correlation analysis of FCGR2A with NET formation markers PADI4 across groups. C , single-cell RNA-seq data ( GSE232237 ) showing FCGR2A expression predominantly in macrophages, with the highest expression in ATC. D , correlation analysis of FCGR2A expression with M0 macrophages, NK cells, and B cells examined by Spearman analysis. E , representative figures of IHC analysis validation of FCGR2A, UBE2C, and NUBPL expression in ATC (n = 11), PDTC (n = 7), and PTC (n = 10) tissues. The numbers at the upper left corner of each IHC image (0–3) indicate the immunoreactivity score for the representative case. The IHC scores were fully plotted in the bar plots assessed by pairwise student’s t tests. ns, not significant; ∗ p < 0.05; ∗∗∗ p < 0.001; and ∗∗∗∗ p < 0.0001. ATC, anaplastic thyroid carcinoma; FCGR2A, Fc fragment of IgG receptor IIa; IHC, immunohistochemistry; log 2 FC, log 2 fold change; NK, natural killer; PDTC, poorly differentiated thyroid carcinoma; PTC, papillary thyroid carcinoma; PADI4, peptidyl arginine deiminase 4; NUBPL, nucleotide-binding protein-like; UBE2C, ubiquitin-conjugating enzyme E2 C.

Article Snippet: Primary antibodies—including FCGR2A (RRID: AB_2246912 , Cat.15625-1-AP, Proteintech, 1:600 dilution), ubiquitin-conjugating enzyme E2 C (UBE2C) (RRID: AB_11232220 , Cat.66087-1-Ig, Proteintech, 1:500), and nucleotide-binding protein-like (NUBPL) (RRID: AB_2878402 , Cat.17393-1-AP, Proteintech, 1:200)—were diluted in antibody dilution buffer and applied to tissue sections.

Techniques: Single Cell, RNA Sequencing, Expressing, Biomarker Discovery, Immunohistochemistry, Binding Assay, Ubiquitin Proteomics

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: VitD promotes M2 microglial polarization in LPS-stimulated BV-2 cells and mice. A-B CCK-8 assays of BV-2 cell viability following treatment with various concentrations of VitD, with or without LPS stimulation. Data are representative of three independent experiments and shown as mean ± SD. C Western blot analysis of CD16, CD18 (M1 markers), CD206 and ARG1 (M2 markers) in LPS- or LPS + VitD (VitD)-treated BV-2 cells and primary mouse microglia. Data are representative of three independent experiments and shown as mean ± SD. n = 3 ** P < 0.01. One-way ANOVA and Tukey’s test. D Immunofluorescence analysis of the levels of CD16 and CD206 in LPS- or LPS + VitD (VitD)-treated BV-2 cells. Scale bar: 50 μm. Right panel: Quantification of CD16 and CD206 IF intensity. Six regions were randomly selected for each group, and the IF intensities of CD16 and CD206 were quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. Red arrows indicate CD16-positive microglia and green arrows indicate CD206-positive microglia. E ELISA analysis of supernatant levels of TNF-α, IL1β, IL-6, IL-4 and IL-10 in LPS- or LPS + VitD (VitD)-treated BV-2 cells. n = 6. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. F Immunofluorescence analysis of the levels of CD16 and CD206 in cerebral cortex and hippocampal CA1 area of LPS- or LPS + VitD(VitD)-treated mice. Scale bar: 20 μm. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. Red arrows indicate CD16-positive microglia and green arrows indicate CD206-positive microglia.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: CCK-8 Assay, Western Blot, Immunofluorescence, Software, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: Mxd1 is essential for VitD-induced microglial M2 polarization in LPS-stimulated conditions. A Immunohistochemistry analysis of Mxd1 expression in cerebral cortex and hippocampal CA1 area hippocampal CA1 area of mice receiving LPS or LPS + VitD (VitD) treatment. Scale bar: 20 μm. Quantification of Mxd1-positive area is shown in the right panel. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot analysis of the protein levels of Mxd1 and c-MYC in BV-2 cells treated with LPS or LPS + VitD (VitD). Data are representative of three independent experiments and shown as mean ± SD. n = 3. ** P < 0.01. One-way ANOVA and Tukey’s test. C Immunofluorescence analysis of the levels of Mxd1 in in BV-2 cells treated with LPS or LPS + VitD (VitD). Scale bar: 100 μm. Right panel: Quantification of Mxd1 IF intensity. Six regions were randomly selected for each group, and the IF intensity of Mxd1 was quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. D Western blot analysis of the protein levels of CD16, CD18, CD206, ARG1 and Mxd1 in LPS or LPS + VitD (VitD)-treated BV-2 cells with either negative control siRNA (siNC), or Mxd1 siRNA (siMxd1). Data are representative of three independent experiments and shown as mean ± SD. n = 3. ns: not significant; * P < 0.05, ** P < 0.01. Two-way ANOVA and Tukey’s test. E ELISA analysis of supernatant levels of TNF-α, IL1β, IL-6, IL-4 and IL-10 in LPS or VitD-treated BV-2 cells with siNC or siMxd1. Data are representative of three independent experiments and shown as mean ± SD. n = 3 ** P < 0.01. Two-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Immunohistochemistry, Expressing, Western Blot, Immunofluorescence, Software, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: Effect of FTO on VitD-induced microglial M2 polarization. A Global m 6 A RNA methylation analysis of total mRNAs in LPS- and LPS + VitD(VitD)-treated BV-2 cells. Data are representative of three independent experiments and shown as mean ± SD. * P < 0.05; ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot of m 6 A methylation regulator (METTL3, METTL14, WTAP, ALKBH5, and FTO) in the same treatment conditions. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. One-way ANOVA and Tukey’s test. C m 6 A quantification in BV-2 cells transfected with negative control siRNA (siNC) or FTO siRNA (siFTO) and treated with LPS or LPS + VitD (VitD). Data are representative of three independent experiments and shown as mean ± SD. * P < 0.05; ** P < 0.01. Two-way ANOVA and Tukey’s test. D Western blot of CD16, CD18, CD206, ARG1 and Mxd1 in the same treatment conditions. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; * P < 0.05; ** P < 0.01. Two-way ANOVA and Tukey’s test. E ELISA of cytokine levels in corresponding supernatants. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. F Immunohistochemistry for Iba-1 in the cerebral cortex and hippocampal CA1 area of mice treated with vehicle, LPS, LPS + VitD, and LPS + VitD + FB23-2. Scale bar: 20 μm. Quantification data are shown in the right panel. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Methylation, Western Blot, Transfection, Negative Control, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: PGC-1α is required for VitD-induced M2 microglial polarization. A Immunofluorescence analysis of PGC-1α nuclear localization in LPS- or LPS + VitD-treated BV-2 cells. Scale bar: 100 μm. Right panel: Quantification of PGC-1α IF intensity. Six regions were randomly selected for each group, and the IF intensity of PGC-1α was quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot analysis of CD16, CD18, CD206 and ARG1 in LPS- or LPS + VitD(VitD)-treated BV-2 cells with vehicle or SR-18,292. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. C ELISA quantification of TNF-α, IL1β, IL-6, IL-4 and IL-10 in culture supernatants from the same experimental groups. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. D Immunohistochemical staining and quantification of Iba-1-positive cells in cerebral cortex and hippocampal CA1 area of LPS- or VitD-treated mice with vehicle or SR-18,292, and the quantification of Iba-1 positive cells in each group was shown in the low panel. Scale bar: 20 μm. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Immunofluorescence, Software, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining