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(A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using <t>anti-CD11b,</t> anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.
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(A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using <t>anti-CD11b,</t> anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.
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(A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using <t>anti-CD11b,</t> anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.
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(A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using <t>anti-CD11b,</t> anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.
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(A) Gating strategy for sorting singlets of B and T cells. (B) Sorted counts. Leukocytes were isolated from previously frozen blood clots and dissociated by following section A of this protocol. Cells were stained with an antibody panel targeting CD3, CD19, CD11b, and <t>CD11c.</t> Viability marker was not used due to interest in nuclei from subpopulations of non-viable leukocytes. After doublet exclusion and gating on height and area of cells, T-cell subsets were identified as CD3+ populations. B-cell subsets were identified as CD19+ populations. CD11b and CD11c were used to exclude monocytes. Single-stain controls were used for compensation. During this sort, we successfully sorted 57,894 T cells and 24,648 B cells. The full report is available in Supplementary material (Report S1).
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Image Search Results


(A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using anti-CD11b, anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: Bloodstream-associated Salmonella Typhimurium and Enteritidis iNTS pathovariants hyper-replicate in human macrophages

doi: 10.64898/2026.01.28.702237

Figure Lengend Snippet: (A) Representative phase contrast images of THP-1 cells untreated versus THP-1 cells differentiated with Condition A (PMA 10 ng/mL for 24 h), Condition B (PMA 10 ng/mL for 24 h followed by a 24 h recovery in fresh media), Condition C (PMA 10 ng/mL for 48 h) and Condition D (PMA 50 ng/mL for 48 h). (B) Representative flow cytometric analysis of THP-1 cells and macrophage-induced cells stained using anti-CD11b, anti-CD14 and anti-CD36 antibodies. (C) Median fluorescence intensity (MFI) of CD11b, CD14 and CD36. Data are displayed as mean ± SD from n ≥ 3. Statistical analyses performed using the Brown-Forsythe and Welch ANOVA test and the Dunnet T3 multiple comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Macrophage surface markers expression was assessed via flow cytometry by staining the cells with anti CD11b (FITC, #130-110-552, Miltenyi Biotec), anti CD14 (APC, #130-110-520, Miltenyi Biotec), anti CD36 (APC-Vio770, #130-110-743, Miltenyi Biotec), anti CD86 (PerCP-Vio700, #130-116-267, Miltenyi Biotec), anti HLA-DR (VioGreen, #130-111-795, Miltenyi Biotec), anti CD163 (PE, #130-112-128, Miltenyi Biotec), anti CD206 (VioBlue, #130-127-809, Miltenyi Biotec) according to manufacturer’s instructions.

Techniques: Staining, Fluorescence, Comparison

(A) Gating strategy for sorting singlets of B and T cells. (B) Sorted counts. Leukocytes were isolated from previously frozen blood clots and dissociated by following section A of this protocol. Cells were stained with an antibody panel targeting CD3, CD19, CD11b, and CD11c. Viability marker was not used due to interest in nuclei from subpopulations of non-viable leukocytes. After doublet exclusion and gating on height and area of cells, T-cell subsets were identified as CD3+ populations. B-cell subsets were identified as CD19+ populations. CD11b and CD11c were used to exclude monocytes. Single-stain controls were used for compensation. During this sort, we successfully sorted 57,894 T cells and 24,648 B cells. The full report is available in Supplementary material (Report S1).

Journal: Bio-protocol

Article Title: Nuclei Isolation Methods on Frozen Clotted Blood Samples

doi: 10.21769/BioProtoc.5573

Figure Lengend Snippet: (A) Gating strategy for sorting singlets of B and T cells. (B) Sorted counts. Leukocytes were isolated from previously frozen blood clots and dissociated by following section A of this protocol. Cells were stained with an antibody panel targeting CD3, CD19, CD11b, and CD11c. Viability marker was not used due to interest in nuclei from subpopulations of non-viable leukocytes. After doublet exclusion and gating on height and area of cells, T-cell subsets were identified as CD3+ populations. B-cell subsets were identified as CD19+ populations. CD11b and CD11c were used to exclude monocytes. Single-stain controls were used for compensation. During this sort, we successfully sorted 57,894 T cells and 24,648 B cells. The full report is available in Supplementary material (Report S1).

Article Snippet: Flow cytometry antibodies (only applicable if flow sorting) a. FITC CD3 antibody (Miltenyi Biotec, catalog number: 130-113-700) b. PE CD19 antibody (Miltenyi Biotec, catalog number: 130-114-172) c. VioBlue CD11B (Miltenyi Biotec, catalog number: 130-110-616) d. APC CD11C (Miltenyi Biotec, catalog number: 130-114-110) e. Other antibodies (optional): Add other suitable antibodies to tailor the cell population of interest 3.

Techniques: Isolation, Staining, Marker