cd11b surface marker  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: BCLAF1 links RNA splicing to ATF4-dependent metabolic adaptation in acute myeloid leukemia

doi: 10.64898/2026.01.19.700325

Figure Lengend Snippet: A. BCLAF1 expression levels across cancer types from the TCGA dataset. B. BCLAF1 expression in LAML patient samples (n = 173) and normal hematopoietic tissues (n = 70), shown as log 2 (TPM + 1). C. Growth competition experiments showing the percentage of GFP-positive cells between day 2 and day 15 following infection with LentiCRISPRv2GFP constructs expressing individual sgRNAs targeting the indicated genes. Data represent mean ± SEM of three biological replicates (*p < 0.05, ***p < 0.001; two-way ANOVA). D. Immunoblot analysis of BCLAF1 protein levels in shCONTROL and shBCLAF1 cells treated or not with DOX for 96 hours. β-tubulin was used as a loading control. E. Flow cytometry analysis showing the percentage of CD11b-positive cells in shCONTROL and shBCLAF1 cells treated with DOX for 96h. Data represent mean ± SEM of three biological replicates (*p < 0.05, ***p < 0.001; unpaired t-test). F. Dot plot of GO terms enriched among genes up-regulated in BCLAF1-low versus BCLAF1-high patient samples from the BEAT-AML cohort. GeneRatio indicates the proportion of significantly upregulated genes associated with each GO term. Dot size reflects the number of genes per term; color denotes adjusted p-value. G. Kaplan-Meier curve shows survival of mice transplanted with MLL-AF9 expressing Bclaf1 f/f (black) or Vav-Cre:Bclaf1 f/f (blue) HSPCs. Briefly, HSPCs from CD45.2 + Bclaf1 f/f or Vav-Cre:Bclaf1 f/f mice were transduced with retrovirus co-expressing MLL-AF9 and GFP then transplanted with 300,000 CD45.1 + recipient bone marrow cells into lethally-irradiated CD45.1 + recipient mice (schematic in Figure S1G). Leukemic burden was assessed by percentage of GFP + cells in peripheral blood. Mice were euthanized once peripheral blood had ≥ 80% leukemia. Inset shows median survival. Bclaf1 f/f , n=23; Vav-Cre:Bclaf1 f/f , n=24. H. Primary Bclaf1 f/f AMLs isolated from mice in G were transduced with retroviral vector expressing Thy1.1 (empty control, black) or Thy1.1 and Cre recombinase (blue) then expanded as mixed co-cultures with nontransduced cells. Bar graphs show percentage of transduced cells (Thy1.1 + ) at day 7 of mixed co-culture relative to day 0 transduction efficiency. Data repesent mean ± SD of 3 independent experiments (**p ≤ 0.01; unpaired t-test). I. Primary Bclaf1 f/f AMLs isolated from mice in G were transduced with retroviral vectors as in D. Thy1.1 + cells were enriched by flow cytometric sorting and cultured in complete methocult media. Total colony forming units were quantitated at day 7 of culture. Data repesent mean ± SD of 3 independent experiments (*p ≤ 0.05; unpaired t-test).

Article Snippet: These samples were stained using CD11b surface marker and Annexin V kit (Annexin V-FITC Kit, Miltenyi Biotec).

Techniques: Expressing, Infection, Construct, Western Blot, Control, Flow Cytometry, Transduction, Irradiation, Isolation, Retroviral, Plasmid Preparation, Co-Culture Assay, Cell Culture

Journal: bioRxiv

Article Title: BCLAF1 links RNA splicing to ATF4-dependent metabolic adaptation in acute myeloid leukemia

doi: 10.64898/2026.01.19.700325

Figure Lengend Snippet: A. Immunoblot analysis of BCLAF1 and ATF4 protein levels in shCONTROL and shBCLAF1 cells treated with DOX for 96 hours. β-tubulin was used as a loading control. B. Immunoblot analysis of BCLAF1 and ATF4 protein levels in MLL-AF9-expressing AML cells of indicated genotypes. GAPDH was used as a loading control. C. Immunoblot analysis of BCLAF1 and ATF4 protein levels in shCONTROL and shATF4 cells treated with DOX for 96 hours. β-tubulin was used as a loading control. D. Cell proliferation assay (CellTiter-Blue-based) in shCONTROL and shATF4 cells treated or not with DOX for 96 hours, followed by 96 additional hours in culture. Data represent mean ± SEM of three biological replicates (*p < 0.01; paired t-test). E. Flow cytometry analysis showing the percentage of CD11b-positive cells in shCONTROL and shATF4 cells treated with DOX for 96h. Data represent mean ± SEM of four biological replicates (***p < 0.001; unpaired t-test). F. Annexin V/PI flow cytometry analysis showing the distribution of live, early apoptotic, late apoptotic, and necrotic cells following 96h of DOX treatment. Data represent mean ± SEM of three biological replicates (two-way ANOVA). G. RT-qPCR experiments showing relative mRNA levels of the indicated transcripts in shCONTROL and shATF4 cells treated with DOX for 96 hours. Data were normalized to GAPDH and represent mean ± SEM of three biological replicates (**p < 0.01; unpaired t-test). H. Immunoblot analysis of ATF4, SLC7A5, ASS1, PHGDH and PYCR1 protein levels in shCONTROL and shATF4 cells treated or not with DOX for 96 hours. β-tubulin was used as a loading control.

Article Snippet: These samples were stained using CD11b surface marker and Annexin V kit (Annexin V-FITC Kit, Miltenyi Biotec).

Techniques: Western Blot, Control, Expressing, Proliferation Assay, Flow Cytometry, Quantitative RT-PCR

Journal: Journal of Orthopaedic Translation

Article Title: An aptamer specifically targeting mCRP ameliorates experimental arthritis

doi: 10.1016/j.jot.2025.08.010

Figure Lengend Snippet: Development of CAIA in hepatocyte-specific CRP knockout mice. (A) Schematic diagram illustrating the development of CAIA in CRP +/+ and CRP ΔHep/ΔHep mice. (B) Arthritis scoring of CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction. Mice without CAIA served as non-immunized (NI) controls. (C and D) Serum IL-6 (C) and IL-1β (D) levels in CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction, as determined by ELISA. (E) Photographs of hind paws of CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction. (F) H&E staining of paw sections of CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction. Scale bar = 300 μm. (G) SO&FG staining of paw sections of CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction. Scale bar = 300 μm. (H) IF staining of paw sections of CRP +/+ and CRP ΔHep/ΔHep mice with or without CAIA induction, using an anti-Vimentin, an anti-CD86, or an anti-MMP-13 antibody. Scale bar = 50 μm. (I – K) Quantification of expression of Vimentin (I) , CD86 (J), and MMP-13 (K) on IF-stained sections. Data are represented as mean ± SD of n = 5 per group. P-values from one-way ANOVA (C, D, I-K) or two-way ANOVA (B): ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Surface marker CD86 was stained directly with anti-human CD86 antibody (Proteintech, Cat# 13395-1-AP, CN).

Techniques: Knock-Out, Enzyme-linked Immunosorbent Assay, Staining, Expressing

Journal: Journal of Orthopaedic Translation

Article Title: An aptamer specifically targeting mCRP ameliorates experimental arthritis

doi: 10.1016/j.jot.2025.08.010

Figure Lengend Snippet: Establishment of surgical DMM in hepatocyte-specific CRP knockout mice. (A) Schematic diagram illustrating the establishment of surgical DMM in CRP +/+ and CRP ΔHep/ΔHep mice. (B) μCT scans of knee joints of CRP +/+ and CRP ΔHep/ΔHep mice after sham or DMM surgery, with black arrows indicating the osteophyte formation. Scale bar = 1 mm. (C) H&E staining of joint sections of CRP +/+ and CRP ΔHep/ΔHep mice after sham or DMM surgery. Scale bar = 300 μm. (D) SO&FG staining of joint sections of CRP +/+ and CRP ΔHep/ΔHep mice after sham or DMM surgery. Scale bar = 300 μm. (E) IF staining of joint sections of CRP +/+ and CRP ΔHep/ΔHep mice after sham or DMM surgery, using an anti-vimentin, an anti-CD86, or an anti-COL2A1 antibody. Scale bar = 50 μm. (F – I) Quantification of OARSI score (F) , cartilage area (G) , osteophyte score (H), and synovitis score (I) of CRP +/+ and CRP ΔHep/ΔHep mice after sham or DMM surgery. (J – L) Quantification of expression of vimentin (J) , CD86 (K), and COL2A1 (L) on IF-stained sections. Data are represented as mean ± SD of n = 5 per group. P-values from one-way ANOVA (F–L): ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Surface marker CD86 was stained directly with anti-human CD86 antibody (Proteintech, Cat# 13395-1-AP, CN).

Techniques: Knock-Out, Staining, Expressing

Journal: Journal of Orthopaedic Translation

Article Title: An aptamer specifically targeting mCRP ameliorates experimental arthritis

doi: 10.1016/j.jot.2025.08.010

Figure Lengend Snippet: Therapeutic effects of ApmCRP3 in CIA mice. (A) Schematic diagram of experimental design using CIA mice. Briefly, CIA mice were treated with PBS, 1,6-bisPC, NC, or ApmCRP3 every 3 days for 45 days. NC or ApmCRP3 was intravenously injected at a dose of 25 mg/kg, and 1,6-bisPC was intraperitoneally administered at a dose of 3 mg/kg. (B) Arthritis scoring of NI or CIA mice from different treatment groups. (C) μCT scans of hind paws of NI or CIA mice from different treatment groups, with black arrows indicating the bone erosion. (D) H&E staining of paw sections of NI or CIA mice from different treatment groups. Scale bar = 300 μm. (E) SO&FG staining of paw sections of NI or CIA mice from different treatment groups. Scale bar = 300 μm. (F) IF staining of paw sections of NI or CIA mice from different treatment groups, using an anti-vimentin, an anti-CD86, or an anti-MMP-13 antibody. Scale bar = 50 μm. (G – I) Quantification of expression of Vimentin (G) , CD86 (H) , and MMP-13 (I) on IF-stained sections. Data were represented as mean ± SD of n = 6 per group. P-values from one-way ANOVA (G–I) or two-way ANOVA (B): ∗p < 0.05, ∗∗∗p < 0.001.

Article Snippet: Surface marker CD86 was stained directly with anti-human CD86 antibody (Proteintech, Cat# 13395-1-AP, CN).

Techniques: Injection, Staining, Expressing

Journal: Journal of Orthopaedic Translation

Article Title: An aptamer specifically targeting mCRP ameliorates experimental arthritis

doi: 10.1016/j.jot.2025.08.010

Figure Lengend Snippet: Therapeutic effects of ApmCRP3 in DMM mice. (A) Schematic diagram of experimental design using DMM mice. Briefly, DMM mice were treated with PBS, 1,6-bisPC, NC, or ApmCRP3 every 3 days for 49 days. NC or ApmCRP3 was intravenously injected at a dose of 25 mg/kg, and 1,6-bisPC was intraperitoneally administered at a dose of 3 mg/kg. (B) μCT scans of knee joints of sham or DMM mice from different treatment groups, with black arrows indicating the osteophyte formation. (C) H&E staining of joint sections of sham or DMM mice from different treatment groups. Scale bar = 300 μm. (D) SO&FG staining of joint sections of sham or DMM mice from different treatment groups. Scale bar = 300 μm. (E) IF staining of joint sections of sham or DMM mice from different treatment groups, using an anti-vimentin, an anti-CD86, or an anti-COL2A1 antibody. Scale bar = 50 μm. (F – I) Quantification of OARSI score (F) , cartilage area (G) , osteophyte score (H), and synovitis score (I) of sham or DMM mice from different treatment groups. (J – L) Quantification of expression of vimentin (J) , CD86 (K), and COL2A1 (L) on IF-stained sections. Data were represented as mean ± SD of n = 6 per group. P-values from one-way ANOVA (F–L): ∗∗∗p < 0.001.

Article Snippet: Surface marker CD86 was stained directly with anti-human CD86 antibody (Proteintech, Cat# 13395-1-AP, CN).

Techniques: Injection, Staining, Expressing