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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with <t>the</t> <t>anti-RFP</t> marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with the anti-RFP marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Journal: Materials Today Bio

Article Title: Electrical stimulation and stem cell subdural implantation decrease microglia reactivity after spinal cord injury

doi: 10.1016/j.mtbio.2026.103329

Figure Lengend Snippet: EPB implantation and ES effects on the implanted hMSCs and iNSCs . A) Representative schematic drawing illustrating the sampling method applied. Spinal cords were dissected, sagittally cryo-sectioned and immunostained for further evaluations after 16 or 46 DPI (short or long term). B, C) Immunofluorescence images and relative magnifications of sagittal spinal cord sections of Sham and Stimulated SCI rats stained with specific marker of human antigen, STEM121 (green), with specific marker of MSCs, CD105 (red), with specific marker of NSCs, SOX2 (magenta), and the nuclei (DAPI, blue) marker at 16 ( B ) and 46 ( C ) DPI. Total implanted cells 1.3 × 10 6 (0.8 × 10 6 for MSC and 0.5 × 10 6 for iNSC). White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 100 μm (scale bar of the magnification = 10 μm and 50 μm respectively). D-F) Histograms of the percentage of ( D ) STEM121+ cells (total implanted cells), ( E ) CD105+/STEM121+ cells (all the implanted MSCs) and ( F ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). G-I) Histograms of the percentage of ( G ) STEM121+ cells (total implanted cells), ( H ) CD105+/STEM121+ cells (all the implanted MSCs) and ( I ) SOX2+/STEM121+ cells (all the implanted iNSCs) in the parenchyma of Sham and Stimulated SCI rats at 46 DPI (Sham n = 9, Stim n = 10). J) Example immunofluorescence images and relative magnification of sagittal spinal cord section stained with specific marker of differentiating neurons, β3Tubulin (red), with the anti-GFP marker for the implanted hMSCs (green), with the anti-RFP marker for the implanted iNCSs (yellow) and the nuclei (DAPI, blue) marker at 16 DPI. White asterisk indicates the lesion site; White arrows indicate hMSCs; yellow arrow indicates iNSCs. Scale bar = 500 μm (scale bar of the magnifications꓿100 μm). K) Histogram of the percentage of β3Tubulin + hMSCs and β3tubulin + iNSCs in the injured spinal cord parenchyma in Sham and Stimulated SCI rats at 16 DPI (Sham n = 6, Stim n = 5). Data are expressed as mean ± SEM. Statistical differences were evaluated by unpaired Student t -test, ∗p < 0.05, ∗∗p < 0.01. Abbreviations: B3Tub: β III Tubulin; CD105: cluster of differentiation 105; DAPI: 4′,6-diamidino-2-phenylindole, dihydrochloride; DPI: days post implantation; GFP: green fluorescent protein; hMSCs: mesenchymal stem cells; ns; not significant; iNSCs: induced neuronal stem cells; RFP: red fluorescent protein; SCI: spinal cord injured. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: guinea pig anti-RFP , 1:400 , 390004, Synaptic System.

Techniques: Sampling, Immunofluorescence, Staining, Marker