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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
Premium Density Gradient Centrifugation, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by <t>Ficoll</t> gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) <t>Flow</t> <t>cytometric</t> histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.
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ficoll-paque premium 1.084  (Aladdin Scientific Corporation)
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DescriptionFicoll-Paque PREMIUM products are a range of sterile, ready-to-use density gradient media for the preparation of mononuclear cells.Ficoll-Paque PREMIUM products are based on Ficoll-Paque PLUS (see Ficoll-Paque PLUS) which has been used for more than
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ficoll-paque premium  (Aladdin Scientific Corporation)
N/A
DescriptionFicoll-Paque PREMIUM products are a range of sterile, ready-to-use density gradient media for the preparation of mononuclear cells.Ficoll-Paque PREMIUM products are based on Ficoll-Paque PLUS (see Ficoll-Paque PLUS) which has been used for more than
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SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by Ficoll gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) Flow cytometric histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.

Journal: Disease Models & Mechanisms

Article Title: Infiltrating circulating monocytes provide an important source of BMP4 at the early stage of spinal cord injury

doi: 10.1242/dmm.049856

Figure Lengend Snippet: SCI induces BMP4 upregulation in circulating monocytes. Blood was collected from SCI and sham rats for ELISA and cytometry analysis. (A,B) ELISA was conducted to detect the BMP4 (A) and noggin (B) levels in the plasma at dpi-1, dpi-7 and dpi-14. (C-K) Peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood by Ficoll gradient separation and analyzed by flow cytometry at dpi-3. (C) The steps and gating strategies of rat circulating monocytes (CD45 + CD11b + ) from the PBMCs. (D,E) Representative flow cytometry plots (D) and statistical data (E) showing the percentage of BMP4 + cells in circulating monocytes. (F,G) Flow cytometric histogram (F) and statistical data (G) showing the mean fluorescence intensity (MFI) of BMP4 expression in circulating monocytes. (H,I) Representative flow cytometry plots (H) and statistical data (I) showing the percentage of BMP4 + cells out of CD86 + monocytes. (J,K) Flow cytometric histogram (J) and statistical data (K) showing BMP4 MFI in CD86 + monocytes. Unpaired two-tailed Student's t -tests (E,G,I,K) and Kruskal–Wallis test followed by Dunn's multiple tests (A,B) were performed. * P <0.05, ** P <0.01 and *** P <0.001. ns, not significant. FSC, forward scatter; SSC, side scatter.

Article Snippet: To prepare peripheral blood mononuclear cells (PBMCs) for flow cytometric detection, whole blood was collected, and PBMCs were separated by Ficoll (17-5446-02, GE Healthcare, Marlborough, MA, USA) density gradient centrifugation according to our previous research ( ).

Techniques: Enzyme-linked Immunosorbent Assay, Cytometry, Isolation, Flow Cytometry, Fluorescence, Expressing, Two Tailed Test