Journal: Experimental and Therapeutic Medicine

Article Title: Appendiceal mucinous tumour resulting in autoamputation of the appendix: A case report and literature review

doi: 10.3892/etm.2025.13043

Figure Lengend Snippet: Immunohistochemical markers in the removed appendix tissue. (A) CKpan positivity; (B) CK7 negativity; (C) CK20 positivity; (D) caudal type homeobox 2 positivity; (E) MUC2 positivity; (F) MUC5AC negativity; (G) PAX-2 negativity; (H) PAX-8 negativity; and (I) Ki-67 negativity. CK, cytokeratin; PAX, paired box; MUC, mucin.

Article Snippet: Primary antibody details were as follows: CK-pan mouse monoclonal antibody (7H8C4; 1:200; cat. no. EM1712-42; HUABIO), CK7 recombinant monoclonal antibody (1:2,000; cat. no. 86153-7-RR; Proteintech Group, Inc.), CK20 recombinant monoclonal antibody (1:4,000; cat. no. 82428-1-RR; Proteintech Group, Inc.), CDX2 recombinant monoclonal antibody (1:1,000; cat. no. 82659-1-RR; Proteintech Group, Inc.), MUC2 polyclonal antibody (1:2,000; cat. no. 27675-1-AP; Proteintech Group, Inc.), MUC5AC polyclonal antibody (1:500; cat. no. 20725-1-AP; Proteintech Group, Inc.), PAX2 polyclonal antibody (1:1,000; cat. no. 29307-1-AP; Proteintech Group, Inc.), PAX8 polyclonal antibody (1:800; cat. no. 10336-1-AP; Proteintech Group, Inc.) and Ki-67 polyclonal antibody (1:4,000; cat. no. 27309-1-AP; Proteintech Group, Inc.).

Techniques: Immunohistochemical staining

Journal: Scientific Reports

Article Title: Regulation of autophagy and its role in late preimplantation during mouse embryo development

doi: 10.1038/s41598-025-11359-2

Figure Lengend Snippet: Rescue of autophagy inhibition by AA supplementation. Developmental rates ( a ) and developmental results ( b ) following culture with 10 μM CQ treatment from the 4/8-cell stages to the blastocyst stage with/without AA supplementation. Scale bars represent 20 μm. ( c ) The embryos at the morula stage, cultured with CQ and AA as shown in panel ‘a,’ were immunostained for Cdx2 and Nanog, as well as with DAPI. Scale bars represent 50 μm. ( d ) The graph shows the numbers of DAPI-positive cells counted in the embryos, as shown in ( c ). Tukey–Kramer’s HSD test, CZB vs. CZB + AA + CQ p = 0.007, CZB + CQ vs. CZB + AA + CQ p = 0.0015. ( e ) The graph shows the proportion of Cdx2- and Nanog-positive cells per total cell number counted in the embryos, as shown in ( c ). Tukey–Kramer’s HSD test, Cdx2: CZB + CQ vs. CZB + AA + CQ p = 0.0012. Nanog: CZB vs. CZ + AA + CQ p = 0.0418, CZB + AA vs. CZB + CQ p = 0.0286, CZB + CQ vs. CZB + AA + CQ p = 0.0005. ( f ) The proportion of positive/negative cells in each embryo, as shown in ( c ). The cells are classified into four types: Nanog-positive (Nanog + , green), Cdx2-positive (Cdx2 + , red), double positive for Nanog and Cdx2 (Nanog + /Cdx2 + , yellow), and double negative (Nanog-/Cdx2-, gray). Tukey–Kramer’s HSD test, Cdx2-/Nanog-: CZB vs. CZB + CQ p = 0.0034, CZB + CQ vs. CZB + AA + CQ p = 0.0045. Cdx2 + /Nanog + : CZB vs. CZB + CQ p = 0.0035. ( g ) The embryos at the morula stage, cultured with CQ and AA as shown in ( a ), were immunostained for TFAP2C and Nanog, as well as with DAPI. Scale bars represent 50 μm. ( h ) The graph shows the proportion of TFAP2C- and Nanog-positive cells per total cell number counted in the embryos, as shown in ( g ). Tukey–Kramer’s HSD test, TFAP2C: no significant differences were observed . Nanog: CZB + CQ vs. CZB + AA + CQ p = 0.0322. ( i ) Relative fluorescence intensity of Nanog (green) and TFAP2C (red) in embryos shown in ( g ). Calibration was performed using DAPI fluorescence intensity.

Article Snippet: The anti-CDX2 monoclonal antibody( 1:500; BioGenex, San Ramon, CA, USA, MU392A-UC) to detect TE cells, anti-Nanog rabbit polyclonal antibody (1:500; Abcam, Cambridge, UK, ab80892) to detect the ICM cells and anti-TFAP2C(1:500; AP-2γ antibody; Santa Cruz Biotechnology, TX, USA, sc-12762) were the primary antibodies used.

Techniques: Inhibition, Cell Culture, Fluorescence

Journal: bioRxiv

Article Title: A Novel Mouse Model Reveals a Role for Mitochondria in Early Lineage Specification and Gastrulation

doi: 10.1101/2025.07.14.664670

Figure Lengend Snippet: A. E3.5 blastocysts from control ( R26R-mitoAtDarT , upper panel) and R26-mitoAtDarT (lower panel) embryos. Immunofluorescence staining shows SOX2 (inner cell mass marker, green), CDX2 (trophectoderm marker, magenta), and nuclei (DAPI, gray). Images were acquired by light-sheet microscopy and analyzed with Imaris software; the Spots module was used for cell quantification. Scale bars, 20 μm. N = 15 embryos per genotype. B. Quantification of total cell number (DAPI+) and lineage-specific cells (SOX2+ and CDX2+) per blastocyst. R26-mitoAtDarT embryos show significantly fewer total cells and reduced numbers of SOX2+ and CDX2+ cells. Notably, the proportion of cells co-expressing SOX2 and CDX2 is increased in R26-mitoAtDarT embryos. C. A stacked bar chart represents the average of the percentage of CDX2+, SOX2+ and CDX2 and SOX2 double positive cells in R26R-mitoAtDarT and R26-mitoAtDarT. The proportion of double-positive cells, relative to the total cell number, was significantly higher for the different markers, P < 0.002.

Article Snippet: Following permeabilization and blocking, embryos were incubated with primary antibodies against CDX2 (Biogenix-MU392A) and SOX2 (Invitrogen-14-9811-82) (1:200 dilution in 10% goat serum, 1% BSA in PBST with 0.3% Triton, blocking buffer) overnight at 4°C for subsequent immunostaining.

Techniques: Control, Immunofluorescence, Staining, Marker, Microscopy, Software, Expressing

Journal: bioRxiv

Article Title: A Novel Mouse Model Reveals a Role for Mitochondria in Early Lineage Specification and Gastrulation

doi: 10.1101/2025.07.14.664670

Figure Lengend Snippet: A. E3.5 blastocysts from control ( R26R-mitoAtDarT , upper panel) and R26-mitoAtDarT (lower panel) embryos. Immunofluorescence staining shows SOX2 (inner cell mass marker, green), CDX2 (trophectoderm marker, magenta), and nuclei (DAPI, gray). Images were acquired by light-sheet microscopy and analyzed with Imaris software; the Spots module was used for cell quantification. Scale bars, 20 μm. N = 15 embryos per genotype. B. Quantification of total cell number (DAPI+) and lineage-specific cells (SOX2+ and CDX2+) per blastocyst. R26-mitoAtDarT embryos show significantly fewer total cells and reduced numbers of SOX2+ and CDX2+ cells. Notably, the proportion of cells co-expressing SOX2 and CDX2 is increased in R26-mitoAtDarT embryos. C. A stacked bar chart represents the average of the percentage of CDX2+, SOX2+ and CDX2 and SOX2 double positive cells in R26R-mitoAtDarT and R26-mitoAtDarT. The proportion of double-positive cells, relative to the total cell number, was significantly higher for the different markers, P < 0.002.

Article Snippet: Cryosections of post-implantation embryos were washed with 0.3% PBST three times for 20 minutes each to ensure permeabilization and complete removal of the OCT. Tissues were then blocked in 10% heat-inactivated goat serum (Himedia) and 1% BSA (Sigma) in 0.3 % PBST for 1 hour, followed by overnight incubation with primary antibodies: CdX2 (Biogenix-MU392A), SOX2 (Invitrogen-14-9811-82), SOX17 (R&D systems-af1924), Brachyury (Abcam-EPR18113), TOMM20 (Thermofisher-MA532148), FLAG-tag (Sigma-F1804), phospho-Histone H3 (pHH3) (Cell Signalling Technology - 9706S), and Cleaved caspase (Cell Signalling Technology - 9661S) at a 1:200 concentration diluted in the blocking buffer, overnight at 4°C.

Techniques: Control, Immunofluorescence, Staining, Marker, Microscopy, Software, Expressing