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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.
Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


The mRNA expression of ITGAV, FAK, PLC, PKC, p65, ERK, JNK, p38, PI3K, Akt, Bax, Bcl2 , and Caspase 3 in E.tenella host cells.

Journal: Poultry Science

Article Title: Pathogenic mechanism of Eimeria tenella Et MIC2 promotes Eimeria tenella invasion and inhibits host cell apoptosis through binding to the ITGAV receptor

doi: 10.1016/j.psj.2026.106922

Figure Lengend Snippet: The mRNA expression of ITGAV, FAK, PLC, PKC, p65, ERK, JNK, p38, PI3K, Akt, Bax, Bcl2 , and Caspase 3 in E.tenella host cells.

Article Snippet: According to the instructions of the Caspase-3 Enzyme Activity Assay Kit (Beyotime, Shanghai, China), cells from each group were lysed for 15 min on ice using lysis buffer.

Techniques: Expressing

The protein activity changes of ITGAV, FAK, PLC, PKC, p65, ERK, JNK, p38, PI3K, Akt, Bax, Bcl2, and Caspase 3 in E.tenella host cells.

Journal: Poultry Science

Article Title: Pathogenic mechanism of Eimeria tenella Et MIC2 promotes Eimeria tenella invasion and inhibits host cell apoptosis through binding to the ITGAV receptor

doi: 10.1016/j.psj.2026.106922

Figure Lengend Snippet: The protein activity changes of ITGAV, FAK, PLC, PKC, p65, ERK, JNK, p38, PI3K, Akt, Bax, Bcl2, and Caspase 3 in E.tenella host cells.

Article Snippet: According to the instructions of the Caspase-3 Enzyme Activity Assay Kit (Beyotime, Shanghai, China), cells from each group were lysed for 15 min on ice using lysis buffer.

Techniques: Activity Assay

Effect of Et MIC2 on E.tenella infection rate and Caspase 3 Activity through ITGAV.

Journal: Poultry Science

Article Title: Pathogenic mechanism of Eimeria tenella Et MIC2 promotes Eimeria tenella invasion and inhibits host cell apoptosis through binding to the ITGAV receptor

doi: 10.1016/j.psj.2026.106922

Figure Lengend Snippet: Effect of Et MIC2 on E.tenella infection rate and Caspase 3 Activity through ITGAV.

Article Snippet: According to the instructions of the Caspase-3 Enzyme Activity Assay Kit (Beyotime, Shanghai, China), cells from each group were lysed for 15 min on ice using lysis buffer.

Techniques: Infection, Activity Assay

Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Article Snippet: After blocking with 5% non-fat milk, membranes were incubated with anti- Caspase-3 antibody (Cell Signaling Technology, 144220, 1:1000) followed by HRP-conjugated secondary antibody.

Techniques: Passaging, Infection, Virus, Disruption, Transmission Assay, Western Blot, Multiplex Assay, Immunofluorescence, Functional Assay, Quantitative RT-PCR, Expressing

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