gsl i isolectin b4 vector laboratories fl 1208  (Vector Laboratories)

Journal: bioRxiv

Article Title: AP-2α/AP-2β transcription factors are key regulators of epidermal homeostasis

doi: 10.1101/2023.12.03.569763

Figure Lengend Snippet: A. Left: Heatmap shows RNA-seq transcript levels of AP-2, EGFR, and EGF family members in P4 skin and the epidermis of 3-week-old, 6-week-old, and 22-month-old WT mice. Middle: Pie charts show relative contributions of AP-2 family members to overall AP-2 transcript levels in epidermis of 3-week-old, 6-week-old, and 22-month-old WT mice. Immunolabeling of adult mouse back skin shows that AP–2α and AP–2β are located in the nuclei of basal and suprabasal keratinocytes. Isolectin B4-DyLight649 (white). Scale bars, 50 mm. Right: Effects of inactivation of AP–2α/AP–2β in keratinocytes on transcript levels of AP-2, EGFR, and EGF family members in 3-week-old and 6-month-old epidermis. B. Left: Effects of inactivation of AP–2α/AP–2β in keratinocytes on transcript levels of AP-2, EGFR, KCTD1 and KCTD15 (regulators of AP-2 activity), and EGF family members in 3-week-old and 6-month-old epidermis. Right: Values shown for 3-week-old epidermis lacking AP–2α or AP–2β in keratinocytes. Log2FC and adjusted p-values (padj) are shown. C. Immunolabeling for AP–2α and AP–2β shows overlapping expression in adult keratinocytes. Back skin of adult WT mice shown. Yellow arrows: interfollicular epidermis. the dermal papilla is not targeted by K14Cre and AP–2α is maintained in the dermal papilla of K14Cre + Tfap2a fl/fl mice [white arrow] that lack AP–2α immunolabeling in keratinocytes [red arrow]). Inactivation of Kctd1 and Kctd15 function in keratinocytes does not affect nuclear localization of AP–2α and AP–2β. Scale bars, 50 mm. D. Transepidermal water loss (TEWL in g/m 2 h) measurements of back skin in newborn (P0) K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice and control littermates. Mean ± SEM; p-value shown for two-sided t-test. E. Body weight (BW) of male K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice and control littermates at P28-P30. Mean ± SEM; p-value shown for two-sided t-test. F. Gender- and age-matched 1-month-old K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice, K14Cre + Tfap2a fl/fl Tfap2b fl/WT mice, and control littermates shown. Hairs show irregular morphology with kinks, bends, and abnormal hair shaft thickness in P29 K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice (green arrow). Typical zigzag hairs, seen in WT controls (black arrow), are not observed in K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice. G. Top: Immunolabeling for keratin 5 (K5; green) and loricrin (red) in back skin of 1-month-old K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice, K14Cre + Tfap2a fl/fl mice, K14Cre + Tfap2b fl/fl mice, and control mice. Bottom: Immunolabeling for Ki67 (red) and labeling with isolectin B4-FITC (green) in the same specimens. Arrows indicate Ki67 + cells. Scale bars, 50 mm. H. Representative images from 2-month-old ear skin of K14Cre + Tfap2a fl/fl Tfap2b fl/fl mice, K14Cre + Tfap2a fl/fl mice, and control mice. Arrows indicate epidermis. Scale bar, 250 mm. I. P7 and P13 K14Cre + Tfap2a fl/fl mice and control littermates are shown. AP–2α inactivation in keratinocytes results in a postnatal growth retardation, delayed hair growth and abnormal morphology of hairs and whiskers.

Article Snippet: FITC-conjugated or DyLight649-conjugated Griffonia Simplicifolia Lectin I (GSL I) isolectin B4 (Vector Laboratories Cat# FL-1201, RRID:AB_2314663; Cat# DL-1208.5) was used at a dilution of 1:100.

Techniques: RNA Sequencing Assay, Immunolabeling, Activity Assay, Expressing, Control, Labeling