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Comprehensive functional validation of CTT Platform after cryopreservation and simulated transport. A) Schematic illustration of the experimental workflow. Fresh or cryopreserved PM@NSC (at −80 °C or −196 °C for 3 months) were thawed and subjected to a 4-h simulated transport at 4 °C prior to in vitro analysis or in vivo transplantation for SCI repair. B) Representative confocal microscopy images assessing post-thaw cell cytoskeletal integrity of NSCs loaded onto PM. Phalloidin (green) for F-actin; DAPI (blue) for nucleus. Scale bar: 50 μm. C) Western blot bands of Nestin, Sox2, and Ki67 show no significant differences in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. D) Representative confocal microscopy images assessing post-thaw cell viability of NSCs loaded onto PM. Calcein AM (green) for live cells; PI (red) for dead cells. Scale bar: 50 μm. E) Western blot bands of <t>Cleaved-Caspase3,</t> Bcl-2, and Bax protein expression in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. F) Quantitative analysis of cell survival rate of NSC in each group (n = 5). G) Quantitative analysis of Nestin/GAPDH, Ki67/GAPDH, and Sox2/β-Actin ratios in each group (n = 3). H) Quantitative analysis of Cleaved-Caspase3/GAPDH, Bcl-2/GAPDH and Bax/GAPDH ratios in each group (n = 3). I) Representative photographs of rat hindlimb motor functions in each group, 8 weeks after SCI. J) MEP results show variations in latency and amplitude in the left hind leg of each group 56 days after SCI K) H&E staining of gastrocnemius muscles indicated variations in muscle fiber morphology among the groups 56 days after SCI. Scale bar: 200 μm L) Sagittal and axial T2-weighted MRI images of rats in each group 56 days after SCI. M) Footprint analysis with print views, footfall patterns, 3D footprints, and 2D footprints revealing differences in gait patterns among separate groups. N) BBB scores demonstrate comparable locomotor functional recovery across all groups, including the Control group and the Cryopreserved-Transport treated group (n = 5). O) Quantitative analysis of gastrocnemius muscle fiber cross-sectional area, indicating similar muscle functional recovery (n = 5). P) Quantitative analysis of MEP latency and amplitude in the left hind leg in each group (n = 5). Q) Quantitative analysis of T2 density in sagittal and coronal planes in spinal cord lesions among groups (n = 5). R) Quantitative footprint analysis on day 56 post-injury included the maximum footprint intensity, footprint positioning, and the regularity index of the left hindlimb (n = 10). All data are presented as the mean ± SEM. Statistical analysis showed no significant differences (n.s.) among the experimental groups. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
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Comprehensive functional validation of CTT Platform after cryopreservation and simulated transport. A) Schematic illustration of the experimental workflow. Fresh or cryopreserved PM@NSC (at −80 °C or −196 °C for 3 months) were thawed and subjected to a 4-h simulated transport at 4 °C prior to in vitro analysis or in vivo transplantation for SCI repair. B) Representative confocal microscopy images assessing post-thaw cell cytoskeletal integrity of NSCs loaded onto PM. Phalloidin (green) for F-actin; DAPI (blue) for nucleus. Scale bar: 50 μm. C) Western blot bands of Nestin, Sox2, and Ki67 show no significant differences in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. D) Representative confocal microscopy images assessing post-thaw cell viability of NSCs loaded onto PM. Calcein AM (green) for live cells; PI (red) for dead cells. Scale bar: 50 μm. E) Western blot bands of <t>Cleaved-Caspase3,</t> Bcl-2, and Bax protein expression in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. F) Quantitative analysis of cell survival rate of NSC in each group (n = 5). G) Quantitative analysis of Nestin/GAPDH, Ki67/GAPDH, and Sox2/β-Actin ratios in each group (n = 3). H) Quantitative analysis of Cleaved-Caspase3/GAPDH, Bcl-2/GAPDH and Bax/GAPDH ratios in each group (n = 3). I) Representative photographs of rat hindlimb motor functions in each group, 8 weeks after SCI. J) MEP results show variations in latency and amplitude in the left hind leg of each group 56 days after SCI K) H&E staining of gastrocnemius muscles indicated variations in muscle fiber morphology among the groups 56 days after SCI. Scale bar: 200 μm L) Sagittal and axial T2-weighted MRI images of rats in each group 56 days after SCI. M) Footprint analysis with print views, footfall patterns, 3D footprints, and 2D footprints revealing differences in gait patterns among separate groups. N) BBB scores demonstrate comparable locomotor functional recovery across all groups, including the Control group and the Cryopreserved-Transport treated group (n = 5). O) Quantitative analysis of gastrocnemius muscle fiber cross-sectional area, indicating similar muscle functional recovery (n = 5). P) Quantitative analysis of MEP latency and amplitude in the left hind leg in each group (n = 5). Q) Quantitative analysis of T2 density in sagittal and coronal planes in spinal cord lesions among groups (n = 5). R) Quantitative footprint analysis on day 56 post-injury included the maximum footprint intensity, footprint positioning, and the regularity index of the left hindlimb (n = 10). All data are presented as the mean ± SEM. Statistical analysis showed no significant differences (n.s.) among the experimental groups. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
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Regulatory effects of SPDEF expression on apoptosis-related protein expression in triple-negative breast cancer cells. (A) Western blot detection of apoptosis-related protein expression in MAD-MB-468 and MAD-MB-468 cell lines. (B-D) Quantitative analysis bar graphs of pro-apoptotic protein expression: Bax (B), <t>cleaved</t> <t>caspase-3</t> (C), and cleaved caspase-9 (D). (E,F) Quantitative analysis bar graphs of anti-apoptotic protein expression: Bcl-2 (E) and Bcl-xL (F). *, P<0.05; **, P<0.01; ***, P<0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; OE, overexpression; SPDEF, SAM pointed domain-containing ETS transcription factor.
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Image Search Results


Comprehensive functional validation of CTT Platform after cryopreservation and simulated transport. A) Schematic illustration of the experimental workflow. Fresh or cryopreserved PM@NSC (at −80 °C or −196 °C for 3 months) were thawed and subjected to a 4-h simulated transport at 4 °C prior to in vitro analysis or in vivo transplantation for SCI repair. B) Representative confocal microscopy images assessing post-thaw cell cytoskeletal integrity of NSCs loaded onto PM. Phalloidin (green) for F-actin; DAPI (blue) for nucleus. Scale bar: 50 μm. C) Western blot bands of Nestin, Sox2, and Ki67 show no significant differences in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. D) Representative confocal microscopy images assessing post-thaw cell viability of NSCs loaded onto PM. Calcein AM (green) for live cells; PI (red) for dead cells. Scale bar: 50 μm. E) Western blot bands of Cleaved-Caspase3, Bcl-2, and Bax protein expression in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. F) Quantitative analysis of cell survival rate of NSC in each group (n = 5). G) Quantitative analysis of Nestin/GAPDH, Ki67/GAPDH, and Sox2/β-Actin ratios in each group (n = 3). H) Quantitative analysis of Cleaved-Caspase3/GAPDH, Bcl-2/GAPDH and Bax/GAPDH ratios in each group (n = 3). I) Representative photographs of rat hindlimb motor functions in each group, 8 weeks after SCI. J) MEP results show variations in latency and amplitude in the left hind leg of each group 56 days after SCI K) H&E staining of gastrocnemius muscles indicated variations in muscle fiber morphology among the groups 56 days after SCI. Scale bar: 200 μm L) Sagittal and axial T2-weighted MRI images of rats in each group 56 days after SCI. M) Footprint analysis with print views, footfall patterns, 3D footprints, and 2D footprints revealing differences in gait patterns among separate groups. N) BBB scores demonstrate comparable locomotor functional recovery across all groups, including the Control group and the Cryopreserved-Transport treated group (n = 5). O) Quantitative analysis of gastrocnemius muscle fiber cross-sectional area, indicating similar muscle functional recovery (n = 5). P) Quantitative analysis of MEP latency and amplitude in the left hind leg in each group (n = 5). Q) Quantitative analysis of T2 density in sagittal and coronal planes in spinal cord lesions among groups (n = 5). R) Quantitative footprint analysis on day 56 post-injury included the maximum footprint intensity, footprint positioning, and the regularity index of the left hindlimb (n = 10). All data are presented as the mean ± SEM. Statistical analysis showed no significant differences (n.s.) among the experimental groups. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Journal: Bioactive Materials

Article Title: Integrated cryopreservation-thawing-transplantation platform for neural stem cell-based spinal cord injury repair

doi: 10.1016/j.bioactmat.2026.01.024

Figure Lengend Snippet: Comprehensive functional validation of CTT Platform after cryopreservation and simulated transport. A) Schematic illustration of the experimental workflow. Fresh or cryopreserved PM@NSC (at −80 °C or −196 °C for 3 months) were thawed and subjected to a 4-h simulated transport at 4 °C prior to in vitro analysis or in vivo transplantation for SCI repair. B) Representative confocal microscopy images assessing post-thaw cell cytoskeletal integrity of NSCs loaded onto PM. Phalloidin (green) for F-actin; DAPI (blue) for nucleus. Scale bar: 50 μm. C) Western blot bands of Nestin, Sox2, and Ki67 show no significant differences in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. D) Representative confocal microscopy images assessing post-thaw cell viability of NSCs loaded onto PM. Calcein AM (green) for live cells; PI (red) for dead cells. Scale bar: 50 μm. E) Western blot bands of Cleaved-Caspase3, Bcl-2, and Bax protein expression in protein expression levels among control (fresh), cryopreserved (−80 °C/-196 °C), and cryopreservation (−80 °C/-196 °C)-transport groups. F) Quantitative analysis of cell survival rate of NSC in each group (n = 5). G) Quantitative analysis of Nestin/GAPDH, Ki67/GAPDH, and Sox2/β-Actin ratios in each group (n = 3). H) Quantitative analysis of Cleaved-Caspase3/GAPDH, Bcl-2/GAPDH and Bax/GAPDH ratios in each group (n = 3). I) Representative photographs of rat hindlimb motor functions in each group, 8 weeks after SCI. J) MEP results show variations in latency and amplitude in the left hind leg of each group 56 days after SCI K) H&E staining of gastrocnemius muscles indicated variations in muscle fiber morphology among the groups 56 days after SCI. Scale bar: 200 μm L) Sagittal and axial T2-weighted MRI images of rats in each group 56 days after SCI. M) Footprint analysis with print views, footfall patterns, 3D footprints, and 2D footprints revealing differences in gait patterns among separate groups. N) BBB scores demonstrate comparable locomotor functional recovery across all groups, including the Control group and the Cryopreserved-Transport treated group (n = 5). O) Quantitative analysis of gastrocnemius muscle fiber cross-sectional area, indicating similar muscle functional recovery (n = 5). P) Quantitative analysis of MEP latency and amplitude in the left hind leg in each group (n = 5). Q) Quantitative analysis of T2 density in sagittal and coronal planes in spinal cord lesions among groups (n = 5). R) Quantitative footprint analysis on day 56 post-injury included the maximum footprint intensity, footprint positioning, and the regularity index of the left hindlimb (n = 10). All data are presented as the mean ± SEM. Statistical analysis showed no significant differences (n.s.) among the experimental groups. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The primary antibodies used in this research are listed below: CD68 (Abcam, Cambridge, UK), CD206 (Abcam, Cambridge, UK), GFAP (Bioss, Beijing, China), iNOS (Abcam, Cambridge, UK), Tuj-1 (Abcam, Cambridge, UK), NF-200 (Invitrogen, CA, USA), MBP (Abcam, Cambridge, UK), HIF-1α (Abcam, Cambridge, UK), VEGFA (Abcam, Cambridge, UK), P-CaMKII (Abcam, Cambridge, UK), CaMKII (Abcam, Cambridge, UK), P-CREB (Cell Signaling Technology, USA), CREB (Cell Signaling Technology, USA), P-PI3K (Cell Signaling Technology, USA), PI3K (Cell Signaling Technology, USA), P-AKT (Cell Signaling Technology, USA), AKT (Cell Signaling Technology, USA), Cleaved-Caspase3 (Cell Signaling Technology, USA), Bcl-2 (Cell Signaling Technology, USA), Bax (Cell Signaling Technology, USA), GAPDH (Proteintech, IL, USA).

Techniques: Functional Assay, Biomarker Discovery, In Vitro, In Vivo, Transplantation Assay, Confocal Microscopy, Western Blot, Expressing, Control, Staining, Muscles

Regulatory effects of SPDEF expression on apoptosis-related protein expression in triple-negative breast cancer cells. (A) Western blot detection of apoptosis-related protein expression in MAD-MB-468 and MAD-MB-468 cell lines. (B-D) Quantitative analysis bar graphs of pro-apoptotic protein expression: Bax (B), cleaved caspase-3 (C), and cleaved caspase-9 (D). (E,F) Quantitative analysis bar graphs of anti-apoptotic protein expression: Bcl-2 (E) and Bcl-xL (F). *, P<0.05; **, P<0.01; ***, P<0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; OE, overexpression; SPDEF, SAM pointed domain-containing ETS transcription factor.

Journal: Translational Cancer Research

Article Title: SPDEF downregulation promotes tumor aggressiveness and poor prognosis in triple-negative breast cancer

doi: 10.21037/tcr-2026-1-0130

Figure Lengend Snippet: Regulatory effects of SPDEF expression on apoptosis-related protein expression in triple-negative breast cancer cells. (A) Western blot detection of apoptosis-related protein expression in MAD-MB-468 and MAD-MB-468 cell lines. (B-D) Quantitative analysis bar graphs of pro-apoptotic protein expression: Bax (B), cleaved caspase-3 (C), and cleaved caspase-9 (D). (E,F) Quantitative analysis bar graphs of anti-apoptotic protein expression: Bcl-2 (E) and Bcl-xL (F). *, P<0.05; **, P<0.01; ***, P<0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NC, negative control; OE, overexpression; SPDEF, SAM pointed domain-containing ETS transcription factor.

Article Snippet: Antibodies against cleaved caspase-3 and GAPDH (catalog numbers: 68773-1-Ig and 60004-1-Ig) were purchased from Wuhan Sanying Biotechnology Co., Ltd. (Wuhan, China), while the cleaved caspase-9 antibody (catalog number: 20750T) was obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot, Negative Control, Over Expression