Journal: Cell reports

Article Title: Regional Control of Hairless versus Hair-Bearing Skin by Dkk2

doi: 10.1016/j.celrep.2018.11.017

Figure Lengend Snippet: (A) Whole mount in situ hybridization for Dkk2 in wild-type mouse embryo at E14.5 (purple signal) reveals high expression levels in cornea and digits (red arrows) and low levels of expression in trunk and head skin between, but not within, hair follicle placodes (yellow arrow) (n = 6 biological replicates). (B) In situ hybridization for Dkk2 in sectioned wild-type mouse P0 dorsal skin (dark blue signal). (C) Higher magnification view of the area indicated by arrows in (B). Note low level expression in inter-follicular dermis (yellow arrows) (n = 3 biological replicates in B and C). (D and E) Ventral (D) and dorsal (E) views of an E15.5 mouse hind limb subjected to whole mount in situ hybridization for Dkk2 (purple-blue signal). Note high expression in the plantar region of the mouse ventral hind limb at E15.5 (arrow in D) (n = 4). (F) Cryosectioned mouse E15.5 hind limb following whole mount in situ hybridization for Dkk2 (purple signal) (n = 5 biological replicates). (G-I) higher magnification views of areas of (F) outlined with a dashed red box (G), a dashed blue box (H), and a dashed brown box (I). Note specific expression of Dkk2 in plantar dermis (arrows in F and G) compared with adjacent footpad skin and dorsal limb dermis at E15.5; low to undetectable levels of Dkk2 in dorsal limb dermis (H); and stronger signal in tissue located at a deeper level in the limb (H). In the limb digit, Dkk2 expression localizes to ventral but not dorsal dermis; relatively high levels of Dkk2 expression are also observed in nail epithelium and in the developing joint region (I). Dashed black lines in (B), (C), and (G)-(I) represent the dermal-epidermal border. (J) Relative levels of Dkk2, Dkk1, Dkk3, Dkk4, and Sostdc1 mRNA in full thickness dorsal paw skin (blue boxes) and plantar skin (pink boxes) from E18.5 mouse embryos, analyzed by qPCR (n = 5 plantar skin samples and n = 5 dorsal paw skin samples in each experiment). Note that only Dkk2 is specifically enriched in plantar skin. (K) Relative levels of Dkk2 mRNA in full thickness mouse Dkk2+/− control (pink box) and Dkk2−/−mutant (black box) plantar skin at P0, analyzed by qPCR (n = 3 littermate controls and n = 3 mutants). Dkk2 transcript levels are undetectable in Dkk2−/− skin. (L and M) Light microscopy of dorsal (L) and ventral (M) left hind paw of a rabbit embryo at E29. Developing hair follicles are present in the ventral plantar region (red arrow, M), as well as in dorsal skin (blue arrow, L). (N) Histology of embryonic rabbit left hind paw at E29 showing the presence of developing hair follicles in ventral plantar skin (red arrow) as well as dorsal paw skin (blue arrow). (O and P) Higher magnification views of the areas indicated in (N) by blue (O) and red (P) dashed boxes, respectively (5 biological replicates for L-P). (Q) Relative levels of rDkk2 mRNA in full thickness dorsal paw skin (blue box) and plantar skin (pink box)from E29 rabbit embryos, analyzed by qPCR (n = 5 plantar skin samples and n = 5 dorsal paw skin samples). Levels of rDkk2 mRNA are not significantly different between embryonic rabbit dorsal and plantar paw skin. All qPCR assays were performed in triplicate for each sample and data were normalized to Gapdh; mean values ± SD are shown. Statistical significance was calculated using Student’s t test. Scale bars, 100 μm (B); 30 μm (C); 750 μm (D and E); 470 μm (F); 150 μm (G-I, O, and P); 3 mm (L and M); 2 mm (N). See also Figure S2.

Article Snippet: Mouse: Sostdc1 lacZ , mixed C57BL/6 / FVB/N / SJL strain background , Dr. Gabriela Loots, Lawrence Livermore National Laboratory , RRID:MGI:4360517.

Techniques: In Situ Hybridization, Expressing, Control, Mutagenesis, Light Microscopy

Journal: Cell reports

Article Title: Regional Control of Hairless versus Hair-Bearing Skin by Dkk2

doi: 10.1016/j.celrep.2018.11.017

Figure Lengend Snippet: (A and B) X-gal-stained whole mounted control Dkk2+/− Axin2lacZ (A) and Dkk2−/− Axin2laaZ (B) hind paws at E17.5. Note elevated Axin2lacZ Wnt reporter activity (blue signal) in the developing plantar region of the Dkk2−/− mutant (B, arrows). The plantar region is outlined by a dashed white line in (A) and (B) and footpad locations are indicated. (C and D) Cryosections of ventral skin from X-gal stained Dkk2+/− Axin2lacZcontrol littermate (C) and Dkk2−/− Axin2lacZ (D) hind paw whole mounts at E17.5. Wnt reporter expression is elevated in plantar papillary dermis and plantar epidermis in the Dkk2−/− mutant compared with the control (yellow arrows). Signaling levels are similar in mutant and control footpads (red arrows). (E-JꞋ) Hind paw sections from Dkk2+/− control littermate (E, EꞋ, G, GꞋ, I, and IꞋ) and Dkk2−/− (F, FꞋ, H, HꞋ, J, and JꞋ) mice at E16.5 (E-FꞋ) or P1 (G-JꞋ) subjected to immunofluorescence (pink signal) for LEF1 (E-HꞋ) or p-catenin (I-JꞋ). Boxed areas in (E)-(J) are shown at higher magnification in (EꞋ)-(JꞋ). Boxed areas in (IꞋ) and (JꞋ) are shown at further increased magnification in the insets. Yellow arrows indicate ectopic expression of LEF1 or β-catenin in Dkk2−/− plantar skin. Green arrow in (JꞋ) inset indicates nuclear and cytoplasmic localization of β-catenin in epithelial cells and white arrow in (JꞋ) inset indicates nuclear localized β-catenin in the dermal condensate of an ectopic hair germ. (K and L) Expression of the Wnt inhibitor Sostdc1 in cryosectioned X-gal stained littermate control (K) and Dkk2−/− mutant (L) ventral paw skin at P1, indicated by expression of a lacZ reporter (blue staining) knocked into the Sostdc1 locus. Note expression of Sostdc1 in plantar and footpad basal epidermis in both control and littermate samples; Sostdc1 also localizes to sweat glands in the footpad of both control and mutant and to ectopic hair follicles in the mutant plantar region. n = 3 mutants and n = 3 littermate controls for each analysis in (A)-(L). EG, eccrine gland; eHFs, ectopic hair follicles. Scale bars, 500 μm (A and B); 300 μm (C and D); 500 μm (E-J); 50 μm (EꞋ-JꞋ); 20 μm (insets in IꞋ and JꞋ); 400 μm (K and L).

Article Snippet: Mouse: Sostdc1 lacZ , mixed C57BL/6 / FVB/N / SJL strain background , Dr. Gabriela Loots, Lawrence Livermore National Laboratory , RRID:MGI:4360517.

Techniques: Staining, Control, Activity Assay, Mutagenesis, Expressing, Immunofluorescence

Journal: Cell reports

Article Title: Regional Control of Hairless versus Hair-Bearing Skin by Dkk2

doi: 10.1016/j.celrep.2018.11.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse: Sostdc1 lacZ , mixed C57BL/6 / FVB/N / SJL strain background , Dr. Gabriela Loots, Lawrence Livermore National Laboratory , RRID:MGI:4360517.

Techniques: SYBR Green Assay, Recombinant, DNA In Situ Hybridization, In Situ Hybridization

Journal: Cell reports

Article Title: In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells

doi: 10.1016/j.celrep.2020.108320

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse: FVB/NJ , The Jackson Laboratory , Cat# JAX:001800, RRID:IMSR_JAX:001800.

Techniques: Virus, Recombinant, Plasmid Preparation, Selection, Isolation, Cell Stimulation, Proliferation Assay, Activation Assay, Software