Journal: Animal Bioscience

Article Title: Transcriptomic analysis identifies CXCL12 as a novel candidate gene for litter size in rabbits

doi: 10.5713/ab.24.0640

Figure Lengend Snippet: CXCL12 overexpression and knockdown in GCs. (A,B) Changes in the expression of CXCL12 after its overexpression and knockdown in GCs. (C,D) The mRNA expression level of PCNA , CCND1 , CDK2 , Bcl-2 and Bax was detected after overexpressing or knocking down CXCL12 in GCs. (E) The protein expression levels of CCND1, PCNA, Bcl-2 and Bax were detected after overexpressing and knocking down CXCL12 in GCs. H, high-litter size group; L, low-itter size group; GAPDH was selected as the internal reference gene (The t test was used for the above analyses comparing two individual samples. * p<0.05; ** p<0.01; *** p<0.001; ns, not significant). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GC, granulosa cells.

Article Snippet: Protein detection was achieved using the following antibodies: anti-CCND1 mouse monoclonal antibody (1:250, Proteintech), anti-PCNA rabbit polyclonal antibody (1:250, Proteintech), anti-Bcl2 rabbit polyclonal antibody (1:250, Proteintech), anti-Bax rabbit polyclonal antibody (1:250, Proteintech), anti-CITED1 rabbit polyclonal antibody (1:50, Proteintech), anti-WNT10B mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-CXCR4 mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-phospho-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam, Cambridge, UK), anti-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam), anti-phospho-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-GAPDH mouse monoclonal antibody (1:2,500, Proteintech), 1:1,000 goat anti-rabbit secondary antibody IgG (Proteintech) and 1:1,000 goat anti-mouse secondary antibody IgG (Proteintech).

Techniques: Over Expression, Knockdown, Expressing

Journal: PLOS One

Article Title: STAG2 mutations in the normal colon induce upregulation of oncogenic pathways in neighbouring wildtype cells

doi: 10.1371/journal.pone.0332499

Figure Lengend Snippet: a. Heatmap depicting the top 20 differentially expressed genes between STAG2 wildtype and mutant populations. b. Using gene set enrichment analysis (GSEA), only the “HALLMARK_TNFA_SIGNALING_VIA_NFKB” gene set was upregulated in wildtype compared to mutant (Normalised enrichment score: 1.75, False discovery rate q -value: 0.078). c. On immunofluorescence staining, we observed increased proliferation of wildtype organoids relative to STAG2 mutants when stained with anti-KI67 antibody. d. Fluorescent intensity of anti-KI67 antibody was statistically significantly upregulated in co-cultured wildtype organoids relative to STAG2 mutant organoids. e. We also observed increased tumorigenicity of wildtype organoids relative to STAG2 mutants when stained with anti-CCND1 antibody. f. Fluorescent intensity of anti-KI67 antibody was statistically significantly upregulated in co-cultured wildtype organoids relative to STAG2 mutant organoids. Scale bars are 50µm. All experiments were performed with N = 3 biological replicates.

Article Snippet: Primary antibodies used included rabbit anti-human STAG2 antibody (1:100, 19837–1-AP, Proteintech, USA), mouse anti-human KI67 antibody (1:500, 66555–6-Ig, Proteintech, USA), mouse anti-human P53 antibody (1:400, 60283–2-Ig, Proteintech, USA), mouse anti-human CCND1 antibody (1:100, 60186–1-Ig, Proteintech, USA), mouse anti-human TERT antibody (1: 100, MA5−16033, Invitrogen, USA), mouse anti-human KRAS antibody (1:250, 415700, Invitrogen, USA), and mouse anti-human TNFα antibody (1:50, MA5−23720, Invitrogen, USA).

Techniques: Mutagenesis, Immunofluorescence, Staining, Cell Culture