monoclonal antibodies sox2  (R&D Systems)

Journal: Nature Communications

Article Title: NR6A1 is essential for neural crest cell specification, formation and survival

doi: 10.1038/s41467-026-68647-2

Figure Lengend Snippet: A In situ hybridization of neuroepithelial markers (purple stain) in null and wildtype littermate control embryos at E9.0. Sox2 , Wnt1 and Pax3 are all expressed in the Nr6a1 null embryos, indicating the neural ectoderm has formed and patterned. B Immunostaining of SOX2 (green), PAX3 (green) and pHH3 (red) on transverse histological sections at E9.0. SOX2 and PAX3 are expressed in the neural plate in both null and control embryos. SOX2 expression is expanded to dorsal regions of the neural plate. PAX3 expression is maintained in the Nr6a1 null, although no PAX3 positive migratory cells could be observed further demonstrating the NCC deficiency. More pHH3 staining can be seen in the neural plate of the Nr6a1 null embryos suggesting the neural ectoderm is maintained in a highly proliferative state. A minimum of 5 embryos were assayed for each in situ hybridization marker and cell proliferation assay.

Article Snippet: The monoclonal antibodies SOX2 (R & D Systems, Minneapolis, MN) and pHH3 (Upstate/Millipore, Billerica, MA) were both used at 1:500.

Techniques: In Situ Hybridization, Staining, Control, Immunostaining, Expressing, Marker, Proliferation Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Fibroblasts fluctuating between mesenchyme and epithelium are involved in hair follicle mesenchyme development

doi: 10.1016/j.bbrep.2025.102006

Figure Lengend Snippet: Transcriptomic characters of K5 lin + fibroblasts in the follicular mesenchyme a The skin of the head, back, and abdomen from Pdgfra-H2B-EGFP; Krt5-Cre; ROSA-CAG-loxP-stop-loxP-tdTomato neonatal mice were harvested. The skin samples were dissociated into single cells, and eGFP+ and tdTomato± cells were sorted by FACS to perform single-cell RNA sequencing. Cells from three mice were pooled. b Quantitative analysis using flow cytometry presents the percentages of eGFP+ and tdTomato± cells in total live cells (2.45 % vs. 49.32 %). c Transcriptomes of fibroblasts were visualized with Uniform Manifold Approximation and Projection (UMAP) and colored according to 10 annotated clusters. d Bubble plots show the representative markers of clusters in ( c ). The circle color and size show average gene expression and percentage of expressing cells in each cluster, respectively. e Percentages of tdTomato± cells by cluster in eGFP+ cells are shown. Fib1(1.70 %) and Fib7 (1.23 %) account for relatively higher proportions than other clusters. f Percentages of tdTomato+ cells in eGFP+ cells are sorted by samples from the head, back, and abdomen. g The analysis of differentially expressed genes between tdTomato-positive and -negative fibroblasts in the Fib1 cluster shows that tdTomato+ cells dominantly expressed representative genes of dermal sheath cells, such as Acta2 , Col11a1 , and Tagln . h Proportion of tdTomato± cells expressing each representative gene for fibroblast subpopulations in dorsal skin fibroblasts are shown. The tdTomato+ cells shared approximately 50 % of the fibroblasts expressing both Sox2 and Acta2 .

Article Snippet: The samples were blocked with 3 % bovine serum albumin (BSA), 0.3 % TritonTMX-100 (Merck), and 0.3 % Polyoxyethylene Sorbitan Monolaurate (Nacalai Tesque) in PBS at 20–25 °C for 1 h. Then, the sections were incubated with rabbit polyclonal anti-mouse keratin 5 (905503, 1:500; BioLegend, San Diego, CA, USA), goat monoclonal anti-mouse SOX2 (AF2018, 5 μg/mL; R&D Systems, Minneapolis, MN, USA), and rabbit monoclonal anti-mouse smooth muscle actin (ab5694, 1:100; Abcam, Cambridge, UK) at 20–25 °C for 18 h. The sections were washed with PBS three times and incubated with Hoechest33342 (1:500; Thermo Fisher Scientific) and Alexa Fluor 647-conjugated goat anti-rabbit IgG (A21245, 1:200; Thermo Fisher Scientific) or Alexa Fluor 647-conjugated donkey anti-goat IgG (A21447, 1:200; Thermo Fisher Scientific) secondary antibodies at 20–25 °C for 4 h. The samples were washed with PBS three times and mounted in ProLong Gold antifade mountant (Thermo Fisher Scientific).

Techniques: RNA Sequencing, Flow Cytometry, Gene Expression, Expressing

Journal: Biochemistry and Biophysics Reports

Article Title: Fibroblasts fluctuating between mesenchyme and epithelium are involved in hair follicle mesenchyme development

doi: 10.1016/j.bbrep.2025.102006

Figure Lengend Snippet: Immunostaining of dorsal skin from Pdgfra-H2B-EGFP; Krt5-Cre; ROSA-CAG-loxP-stop-loxP-tdTomato neonatal and aged mice. a Immunostaining of neonatal dorsal skin shows that both tdTomato- and eGFP- positive cells were observed in dermal papilla, expressing SOX2 (white arrowhead), and dermal sheath, expressing αSMA and CD200 (white arrow). Cells at dermal sheath cup, the edge of dermal papilla, were tdTomato-positive with expressing both SOX2 and αSMA (yellow arrowhead). Krt5 was not currently expressed in tdTomato- and eGFP- double positive cells. Scale bar = 50 μm b TdTomato expressing fibroblasts were located in dermal papilla, dermal sheath, and interstitial dermis (arrow) in aged (49-week-old) mice. Scale bar = 50 μm.

Article Snippet: The samples were blocked with 3 % bovine serum albumin (BSA), 0.3 % TritonTMX-100 (Merck), and 0.3 % Polyoxyethylene Sorbitan Monolaurate (Nacalai Tesque) in PBS at 20–25 °C for 1 h. Then, the sections were incubated with rabbit polyclonal anti-mouse keratin 5 (905503, 1:500; BioLegend, San Diego, CA, USA), goat monoclonal anti-mouse SOX2 (AF2018, 5 μg/mL; R&D Systems, Minneapolis, MN, USA), and rabbit monoclonal anti-mouse smooth muscle actin (ab5694, 1:100; Abcam, Cambridge, UK) at 20–25 °C for 18 h. The sections were washed with PBS three times and incubated with Hoechest33342 (1:500; Thermo Fisher Scientific) and Alexa Fluor 647-conjugated goat anti-rabbit IgG (A21245, 1:200; Thermo Fisher Scientific) or Alexa Fluor 647-conjugated donkey anti-goat IgG (A21447, 1:200; Thermo Fisher Scientific) secondary antibodies at 20–25 °C for 4 h. The samples were washed with PBS three times and mounted in ProLong Gold antifade mountant (Thermo Fisher Scientific).

Techniques: Immunostaining, Expressing

Journal: Biomedicines

Article Title: Morphological and Immunocytochemical Characterization of Paclitaxel-Induced Microcells in Sk-Mel-28 Melanoma Cells

doi: 10.3390/biomedicines12071576

Figure Lengend Snippet: Primary and secondary antibodies.

Article Snippet: SOX2 , Primary antibody: mouse anti-human antibody Mouse monoclonal , , 1:100 , TA302025, Origene, Rockville, MD, USA.

Techniques: Concentration Assay, Recombinant