Journal: Cancers
Article Title: Kallikrein 5 Inhibition by the Lympho-Epithelial Kazal-Type Related Inhibitor Hinders Matriptase-Dependent Carcinogenesis
doi: 10.3390/cancers13174395
Figure Lengend Snippet: Co-expression of LEKTI attenuates matriptase-mediated premalignant skin phenotype. ( A ) The scheme shows the breeding of K5-LEKTI +/0 with K5-Matriptase +/0 mice and the resulting litter of WT, K5-LEKTI +/0 , K5-Matriptase +/0 , and K5-Matriptase +/0 /K5-LEKTI +/0 mice. Images show the outward appearance of these mice at 3 months of age. Matriptase-induced alopecia and ichthyosis are considerably attenuated by co-expression of LEKTI in Matriptase +/0 /K5-LEKTI +/0 . LEKTI +/0 in 3-month-old mice. ( B ) Representative histological appearance of dorsal skin of littermate WT (first column), K5-LEKTI +/0 (second column), K5-Matriptase +/0 (third column), and K5-Matriptase +/0 /K5-LEKTI +/0 mice (forth column) stained by H&E (top panels) and Toluidine Blue (bottom panels) at 3 months of age. Bars = 100 μm. Yellow dashed lines show the limits between epidermis and dermis. Black arrows show metachromatically stained dermal mast cells. ( C ) Quantification of epidermal thickness in littermate WT ( n = 7, black dots), K5-LEKTI +/0 ( n = 6, orange dots), K5-Matriptase +/0 ( n = 5, purple dots), and K5-Matriptase +/0 /K5-LEKTI +/0 ( n = 11, green dots) at 3 months of age. Data are expressed in mean ± SD. ( D ) Quantification of the dermal mast cell accumulation in the skin of littermate WT (black dots), K5-LEKTI +/0 (orange dots), K5-Matriptase +/0 (purple dots), and K5-Matriptase +/0 /K5-LEKTI +/0 (green dots) at 3 months of age. Data are expressed in mean ± SD. ( E – J ) Myeloid cellular infiltration was evaluated in skin samples by flow cytometry. ( E ) Total cells (×10 5 ) were assessed by automated cell counter using trypan blue. The percentage of ( F ) leukocytes (CD45 + ), ( G ) neutrophils (Ly6G + gated on CD45 + ), ( H ) dendritic cells (DC–CD11c + MHCII High gated on CD45 + Ly6G), ( I ) other myeloid cells (MY–CD11c + MHCII + gated on CD45 + Ly6G − ), and ( J ) macrophages (CD64 + gated on myeloid cells) in the skin. WT (black dots), K5-LEKTI +/0 (orange dots), K5-Matriptase +/0 (purple dots), and K5-Matriptase +/0 /K5-LEKTI +/0 (green dots). ( K ) Representative flow cytometry of skin samples by groups. Data are representative of one experiment ( n = 3/group) and are expressed as means ± SD. p -values (One-Way ANOVA with Tukey’s post-hoc test) displayed in the graphs.
Article Snippet: The following conjugated primary antibodies were used for flow cytometry: anti-CD45-PeCy7 (clone 30-F11), anti-Ly6G-APC (clone IA8), anti-CD11c-FITC (clone HL3), anti-MHCII-BB700 (clone M5/114.15.2) from BD Bioscience (Franklin Lakes, NJ, USA), and anti-CD64-APC-Cy7 (clone X54-5/7.1) and anti-CD16/CD32 monoclonal antibody (clone 93) from eBioscience (San Diego, CA, USA).
Techniques: Expressing, Staining, Flow Cytometry