Journal: Frontiers in Cell and Developmental Biology

Article Title: Screening for novel factors involved in mouse early embryonic development using inhibitor libraries

doi: 10.3389/fcell.2025.1643551

Figure Lengend Snippet: Cathepsin D ( Ctsd ) and Cxcr2 genes are expressed in mouse embryo. (A) Expression level of Ctsd, and Cxcr2 by developmental stage in mice. The RNA-seq data from oocyte to morula stage mouse embryo in public database is reanalyzed. Each line indicates the expression level calculated by log2(TPM+1) values. (B) Expression level of CTSD, and CXCR2 by developmental stage in human. The RNA-seq data from oocyte to morula stage mouse embryo in public database is reanalyzed. Each line indicates the expression level calculated by log2(TPM+1) values. (C) Immunofluorescence staining of Ctsd protein (CTSD) from MII stage oocyte to blastocyst stage embryo. Green indicates CTSD, and blue indicates DAPI The right panel shows the result using a negative control antibody instead of the anti-CTSD antibody. Size bars: 100 µm. (D) Immunofluorescence staining of Cxcr2 protein (CXCR2) from MII stage oocyte to blastocyst stage embryo. Green indicates CXCR2, and blue indicates DAPI The right panel shows the result using a negative control antibody instead of the anti-CXCR2 antibody. Size bars: 100 µm.

Article Snippet: For CTSD detection, an anti-CTSD rabbit polyclonal IgG antibody (1:200, 55021-1-AP, Proteintech group Inc., IL, United States), an anti-CXCR2 (1:200, 19538-1-AP, Proteintech) for CXCR2 detection, and as a negative control, a rabbit IgG isotype control antibody (98136-1-RR, Proteintech) was used.

Techniques: Expressing, RNA Sequencing, Immunofluorescence, Staining, Negative Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Screening for novel factors involved in mouse early embryonic development using inhibitor libraries

doi: 10.3389/fcell.2025.1643551

Figure Lengend Snippet: CRISPR-Cas9 mediated knockout of Ctsd genes in mouse embryos impairs preimplantation development. (A) Experimental workflow for CRISPR-Cas9 mediated gene knockout in one-cell stage embryos via electroporation. (B) Schematic of the Ctsd gene structure. Guide RNAs were designed to target the start codon in exon 1, which is shared across all splicing isoforms. (C) DNA sequences of the wild type (upper) and the knockout embryo (lower). Dashes (−) indicate nucleotides missing from the original target sequence. (D) Developmental outcomes of Ctsd knockout embryos. Data are presented as mean ± SD from three independent experiments. Statistical analysis was performed using the t-test. *p < 0.05.

Article Snippet: For CTSD detection, an anti-CTSD rabbit polyclonal IgG antibody (1:200, 55021-1-AP, Proteintech group Inc., IL, United States), an anti-CXCR2 (1:200, 19538-1-AP, Proteintech) for CXCR2 detection, and as a negative control, a rabbit IgG isotype control antibody (98136-1-RR, Proteintech) was used.

Techniques: CRISPR, Knock-Out, Gene Knockout, Electroporation, Sequencing