Journal: Cell Reports Medicine

Article Title: Selective apoptosis of tumor-associated platelets boosts the anti-metastatic potency of PD-1 blockade therapy

doi: 10.1016/j.xcrm.2025.101984

Figure Lengend Snippet:

Article Snippet: PE/Cyanine5 anti-mouse/human CD11b (clone: M1/70) , Elabscience , CatE-AB-F1081G.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software, Staining, Modification, Clinical Proteomics, Membrane

Journal: Advanced Science

Article Title: The KDM6B/SLC10A2 Axis Suppresses MDSCs Recruitment via ERK/AP‐1 Signaling in Colorectal Cancer

doi: 10.1002/advs.202514086

Figure Lengend Snippet: KDM6B deficiency exacerbates CRC progression and promotes an immunosuppressive microenvironment. A) Schematic of the AOM/DSS‐induced CRC model in the Villin Cre ; KDM6B fl/fl group and KDM6B fl/fl group; each genotype was represented by five biological replicates. B) Longitudinal body weight changes. C) Disease activity index (DAI) scores (0–4 scale) during three DSS cycles. D) Macroscopic view of colon tumors at the endpoint. The pathological area and manifestation of tumors in the colon are indicated by arrows. E) Quantification of tumor number (left) and diameter (right). F) H&E staining of colon tissues. Grading of dysplastic lesions (aberrant crypt foci, adenoma, and high‐grade dysplasia). G) Left: Representative immunohistochemical staining of Ki67 in colon crypts. The black arrows indicate Ki67 + proliferating cells. Right: Quantification of Ki67 + cells per crypt. H) Flow cytometry gating strategy for PMN‐MDSCs (CD11b + Ly6G + ) from the colon epithelium, spleen, and bone marrow. I) Quantification of PMN‐MDSC frequencies and correlation between KDM6B expression and MDSC infiltration in human CRC using the TIMER2.0 database. (J) Flow cytometry of CD4 + and CD8 + T cells in the colon epithelium. K) Correlations between KDM6B expression and CD4/CD8 T‐cell markers in human CRC according to the TIMER2.0 database. L) Flow cytometry analysis of M1 (CD86 + ) and M2 (CD206 + ) macrophages in the colon epithelium. M) Representative subcutaneous tumors from the MC38‐vector and MC38‐KDM6B‐OE groups. N) Tumor growth curves and endpoint tumor weights. O) Flow cytometry of tumor‐infiltrating PMN‐MDSCs and CD4 + /CD8 + T cells. P) Flow cytometry of peripheral blood PMN‐MDSCs and CD4 + /CD8 + T cells. Scale bars: 200 µm. Data are pooled from five independent mice in (B–K) and presented as the means ± SEM. Statistical significance was determined by an unpaired, two‐tailed Student's t test. * p < 0.05, ** p < 0.01. (A) was created with bioRender.com, with permission.

Article Snippet: Antibodies against the following proteins were used: CD45 (E‐AB‐F1136E; Elabscience Biotechnology, Wuhan, China), CD11b (E‐AB‐F1081F; Elabscience), Ly6G (E‐AB‐F1108D; Elabscience), Ly6C (E‐AB‐F1121H; Elabscience), CD3 (E‐AB‐F1013C; Elabscience), CD4 (E‐AB‐F1097F; Elabscience), CD8 (E‐AB‐F1104D; Elabscience), F4/80 (E‐AB‐F0995H; Elabscience), CD86 (E‐AB‐F0994D; Elabscience), and CD206 (E‐AB‐F1135C; Elabscience).

Techniques: Activity Assay, Staining, Immunohistochemical staining, Flow Cytometry, Expressing, Plasmid Preparation, Two Tailed Test

Journal: Advanced Science

Article Title: The KDM6B/SLC10A2 Axis Suppresses MDSCs Recruitment via ERK/AP‐1 Signaling in Colorectal Cancer

doi: 10.1002/advs.202514086

Figure Lengend Snippet: KDM6B deficiency promotes MDSCs‐mediated immunosuppression via the AP‐1/CXCL–CXCR2 axis. A) Immunohistochemical (IHC) staining of Gr1 and CXCR2 in colon tissues from Villin Cre ; KDM6B fl/fl mice and KDM6B fl/fl littermate controls after 10 weeks of AOM/DSS treatment. Quantification of positive pixels per field. B) qPCR analysis of the expression of MDSC markers (S100A8, S100A9, Arg1, TGF‐β, and VEGF‐a) in the intestinal epithelium of Villin Cre ; KDM6B fl/fl mice and KDM6B fl/fl mice. C,D) MC38 murine colon cancer cells stably overexpressing KDM6B (OE‐KDM6B) or empty vector control (MCS) were subcutaneously injected into C57BL/6 mice (n = 5 per group). (C) Immunohistochemical (IHC) staining of Gr1 and CXCR2 in subcutaneous tumors derived from MC38‐MCS and MC38‐OE‐KDM6B cells at the endpoint (14 days post‐injection). Representative images are shown. Scale bars: 200 µm. D) qPCR of the expression of MDSCs markers (S100A8, S100A9, Arg1, TGF‐β, and VEGF‐a) in subcutaneous tumors derived from MC38‐MCS and MC38‐OE‐KDM6B cells at the endpoint. E) Transwell migration assays were performed to evaluate the ability of tumor‐conditioned media (TCM) from MC38 cells, including the MC38‐vector (MCS), MC38‐OE‐KDM6B (OE‐KDM6B), MC38‐shNC (NC), and MC38‐shKDM6B (sh‐KDM6B) groups, to recruit MDSCs. Representative images are shown. Each group had three biological replicates. F) Quantification of migrated MDSCs. G,H) qPCR of the expression of MDSC markers (S100A8, S100A9, Arg1, TGF‐β, and VEGF‐a) after TCM treatment. I,J) CXCR2 blockade experiments: (I) A CXCR2 inhibitor (SB265610, 10 µM) was added into tumor‐conditioned media (TCM) from MC38‐NC cells or MC38‐sh‐KDM6B cells, and Transwell assays were performed to evaluate the migration capacity of MDSCs. Representative images are shown. Each group had three biological replicates. (J) The statistical results of the migration cell count. K–M) In vivo CXCR2 inhibition: (K) Subcutaneous tumors were generated in C57BL/6 mice by injection of MC38 cells with stable KDM6B knockdown (sh‐KDM6B) or negative control (NC). Mice were treated daily with the CXCR2 inhibitor SB265610 (2 mg kg −1 day −1 , i.p.) or vehicle control for 15 days (n = 5). (L) Tumor growth curves. (M) Endpoint tumor weights. (N) Flow cytometry of tumor‐infiltrating PMN‐MDSCs (CD11b + Ly6G + Ly6C − ), CD4 + T cells (CD3 + CD4 + ) and CD8 + T cells (CD3 + CD8 + ). (O) Flow cytometry of peripheral blood from the four experimental groups (NC, NC+CXCR2i, sh‐KDM6B, and sh‐KDM6B+CXCR2i) was performed at the endpoint (Day 15). Gating strategy for PMN‐MDSCs (CD11b⁺Ly6G⁺Ly6C − ), CD4 + T cells (CD3 + CD4 + ) and CD8 + T cells (CD3 + CD8 + ). (P) Quantification of MDSCs and T cells in tumors and peripheral blood. (Q) Immunohistochemical (IHC) analysis of CXCR2 and Gr1⁺ cell infiltration in subcutaneous tumors. The sample size for each group was n = 5. The middle line in the box plots represents the median, whereas the whiskers denote the minimum‐to‐maximum range of the data distribution. The error bars show the means ± SEM. Statistical significance was determined by an unpaired, two‐tailed Student's t test. * p < 0.05, ** p < 0.01.

Article Snippet: Antibodies against the following proteins were used: CD45 (E‐AB‐F1136E; Elabscience Biotechnology, Wuhan, China), CD11b (E‐AB‐F1081F; Elabscience), Ly6G (E‐AB‐F1108D; Elabscience), Ly6C (E‐AB‐F1121H; Elabscience), CD3 (E‐AB‐F1013C; Elabscience), CD4 (E‐AB‐F1097F; Elabscience), CD8 (E‐AB‐F1104D; Elabscience), F4/80 (E‐AB‐F0995H; Elabscience), CD86 (E‐AB‐F0994D; Elabscience), and CD206 (E‐AB‐F1135C; Elabscience).

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Stable Transfection, Plasmid Preparation, Control, Injection, Derivative Assay, Migration, Cell Characterization, In Vivo, Inhibition, Generated, Knockdown, Negative Control, Flow Cytometry, Two Tailed Test

Journal: Neural regeneration research

Article Title: Atherosis-associated lnc_000048 activates PKR to enhance STAT1-mediated polarization of THP-1 macrophages to M1 phenotype.

doi: 10.4103/NRR.NRR-D-23-01355

Figure Lengend Snippet: Figure 1 ｜Verification of M0, M1, and M2 macrophages by cell morphology and RNA and protein expressions. (A) Morphology of M0, M1 and M2 macrophages (magnification, 10×, scale bars: 400 µm). (B) Tumor necrosis factor-α (TNF-α), interleukin (IL)-6 (IL-6), and IL-1β mRNA expression in M0 and M1 macrophages detected by real-time fluorescence quantitative PCR (qRT-PCR). (C) The mRNA expression of CD38 and CD80 in M0 and M1 macrophages was measured by qRT-PCR. (D) IL-10, CCL18, and CCL22 mRNA expression in M0 and M2 macrophages detected by qRT-PCR. (E) The expression of CD163 mRNA in M0 and M2 macrophages was measured by qRT-PCR. (F) The proportion of CD11b-labeled M0 macrophages was determined by flow cytometry. (G) The proportion of CD38-labeled and CD80-labeled M1 macrophages was determined by flow cytometry. Data are displayed as mean ± SD (n = 3). *P < 0.05, **P < 0.01 (independent samples t-test).

Article Snippet: PE anti-human CD11b antibody (Elabscience, Wuhan, China, Cat# E-AB-F1146D), PE mouse anti-human CD38 antibody (Becton, Dickinson and Company, Queensbury, NY, USA, Cat# 560981), or PE mouse anti-human CD80 antibody (Becton, Dickinson and Company, Cat# 560925) were added, and the mixture was incubated at 4°C in the dark for 45 minutes.

Techniques: Expressing, Fluorescence, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Labeling, Flow Cytometry

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: The Thyroid Hormone Analog GC-1 Mitigates Acute Lung Injury by Inhibiting M1 Macrophage Polarization.

doi: 10.1002/advs.202401931

Figure Lengend Snippet: Figure 2. GC-1 selectively inhibited the M1 macrophage polarization. A) Representative images of IHC staining for macrophage marker F4/80, M1 macrophage marker CD86, or M2 macrophage marker CD206 in lung sections of LPS-induced ALI model mice (n = 3), Scale bars: 100 and 20 μm (insets). B) Quantitative statistical results of IHC-stained positive cells (n = 3). C) Representative pictures of F4/80, CD86, and CD206 IF staining in lung sections from ALI mice induced by HCl (n = 3), Scale bars: 50 μm. Quantitative statistical results of fluorescence intensity from D) F4/80+CD86+ macrophages and E) F4/80+CD206+ macrophages (n = 3). F) Representative flow cytometry plots showing the proportions of M1 (CD11b+CD86+CD206−) and M2 (CD11b+CD86−CD206+) macrophages (n = 3). THP-1 cells were treated with LPS (100 ng mL−1) alone or in combination with GC-1 (100 nm) for 24 h after PMA induction. G) CD11b+CD86+CD206−M1 macrophages and H) CD11b+CD86−CD206+ M2 macrophages quantitative statistical results (n = 3). I) Immunoblot analysis of lysates from THP-1 cells (n = 3). J) Immunoblot gray values statistical quantification (n = 3). The values are shown as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns = not significant.

Article Snippet: Flow Cytometry: According to the instructions, APC Anti-Human CD206 Antibody (E-AB-F1161E, Elabscience Biotechnology), PE AntiHuman CD86 Antibody (E-AB-F1012D, Elabscience Biotechnology), and FITC Anti-Human CD11b Antibody (E-AB-F1081C, Elabscience Biotechnology) were separately added.

Techniques: Immunohistochemistry, Marker, Staining, Cytometry, Western Blot