Journal: bioRxiv

Article Title: Convergent deployment of ancestral programs during the evolution of mammalian flight membranes

doi: 10.1101/2022.12.06.518272

Figure Lengend Snippet: (A) Heatmap displaying eigengene expression of WGCNA modules across early patagium development. Columns represent individuals, sorted by increasing weight. Rows represent modules, sorted by their correlation with sample age. Cell color represents module eigengene expression. (B-C) Genes differentially expressed between the patagium primordium and dorsal (B) or shoulder skin (C). Labeled genes are those that were both differentially expressed in the patagium primordium relative to comparison skin regions (upregulated shown in green, downregulated shown in purple), and belonged to the enriched Wnt-signaling Gene Ontology term (GO:0030111). (D) In situ hybridizations of Wnt5a, Wnt11 and Dkk2 . (E) Schematic of tissue recombination experiments. (F) Explants stained for KRT4 and DAPI. Asterisk denotes the mesenchymal condensate. (G) Quantification of epidermal thickness in recombination experiments. (H) Hematoxylin-eosin (H&E) and KRT14 stained, bead-implanted explants. Asterisk shows implanted bead. (I) Quantification of epidermal thickness in bead-implanted explants. Statistical significance in panels G (**** P < 0.0001; N = 5) and I (**** P < 0.0001; N = 4) was assessed using a general mixed effects model ANOVA test. Dotted lines in (D) and (H) delineate the dermis-epidermis boundary. Scale bars: 400μm (zoomed out) and 100μm (zoomed in) in (D); 100μm in (F); and 200μm in (H).

Article Snippet: Immunohistochemistry in sections of sugar glider explants, transgenic mouse skin and bat plagiopatagium was performed using anti-KRT14 (BioLegend: 905303; 1:1000), anti-WNT5A (BiossUSA: bs-1948R; 1:100), and anti-Ki67 (Abcam: ab15580; 1:100).

Techniques: Expressing, Labeling, In Situ, Staining

Journal: bioRxiv

Article Title: Convergent deployment of ancestral programs during the evolution of mammalian flight membranes

doi: 10.1101/2022.12.06.518272

Figure Lengend Snippet: (A) DAPI-stained transverse sections showing dermal cell density in transgenic mice and controls. Yellow box denotes zoomed-in region. (B) Quantification of cell density in transgenic mice and controls. (C) Hematoxylin-Eosin (H&E) and KRT14 stained mouse skin from transgenic mice and controls. (D) Quantification of epidermal thickness in transgenic mice and controls. (E) E14.5 mouse, showing the early ear pinna. Dotted line shows the rostro-caudal axis along which sections were taken. (F) ROIs used for quantifying dermal cell density and epidermal thickness. (G) Sections stained with DAPI and KRT14. (H-I) Quantification of cell density (H) and epidermal thickness (I) in E14.5 mouse pinna. (J) In situ hybridization for Wnt5a in E14.5 mouse pinna. Statistical significance in panels (B and D) (N = 4; **** P < 0.001) and in panels (H and I) (**** P < 0.0001; *** P < 0.001; ** P < 0.01; *P < 0.05; N = 3) was assessed using a general mixed effects model ANOVA test. Post-hoc pairwise comparisons indicated that Region 4 was significantly different from other regions. All other comparisons were statistically not significant and are not displayed. Dotted lines in (C) and (J) delineate the dermis-epidermis boundary. Scale bars: 200μm (zoomed out) and 50μm (zoomed in) in (A); 100μm in (C); 200μm in (F); 50μm in (G); 100μm (zoomed out) and 50μm (zoomed in) in (J).

Article Snippet: Immunohistochemistry in sections of sugar glider explants, transgenic mouse skin and bat plagiopatagium was performed using anti-KRT14 (BioLegend: 905303; 1:1000), anti-WNT5A (BiossUSA: bs-1948R; 1:100), and anti-Ki67 (Abcam: ab15580; 1:100).

Techniques: Staining, Transgenic Assay, In Situ Hybridization

Journal: bioRxiv

Article Title: Convergent deployment of ancestral programs during the evolution of mammalian flight membranes

doi: 10.1101/2022.12.06.518272

Figure Lengend Snippet: (A) The plagiopatagium (red arrowheads) of Seba’s short-tailed bat is shown in an adult animal and (B) in embryonic stages. This structure differs from other patagia in the bat wing, such as the chiropatagium (blue arrowheads). (C) Quantification of dermal cell density and epidermal thickness along the dorsal-ventral axis in an embryonic bat (St. 16) within multiple ROIs (highlighted in yellow) and illustrated in a heat map. (D and E) Bat plagiopatagium relative to dorsal skin in transverse sections, with cell density visualized by DAPI (D) and epidermal thickness visualized by IHC for KRT14 (E). (F) Whole-mount in situ hybridization of embryonic bats showing Wnt5a expression in the developing plagiopatagium (red arrowheads) relative to dorsal skin. Wnt5a is also detected in the limbs (black arrowheads) (fl: forelimbs; hl: hindlimbs). (G) Volcano plot of genes differentially expressed between the sugar glider patagium and the neighboring dorsal skin during its outgrowth. Patagium-upregulated genes belonging to limb development Gene Ontology terms are labeled in green. (H) Venn diagram showing genes with known roles in limb outgrowth and patterning convergently-upregulated in the developing lateral patagia of sugar gliders and bats. Photo credit in (A): Andrew Morffew.

Article Snippet: Immunohistochemistry in sections of sugar glider explants, transgenic mouse skin and bat plagiopatagium was performed using anti-KRT14 (BioLegend: 905303; 1:1000), anti-WNT5A (BiossUSA: bs-1948R; 1:100), and anti-Ki67 (Abcam: ab15580; 1:100).

Techniques: In Situ Hybridization, Expressing, Labeling

Journal: Journal of Cancer

Article Title: The role of β-catenin in the initiation and metastasis of TA2 mice spontaneous breast cancer.

doi: 10.7150/jca.19723

Figure Lengend Snippet: Figure 1. A. Morphological changes in different breast lesions in TA2 mice. a) Normal breast mammary gland tissue from TA2 mice without SBC (×100, black arrowheads). b) Ductal hyperplasia of breast mammary gland tissue from TA2 mice with SBC (×200, black arrowheads). c) Atypical ductal hyperplasia of breast mammary gland tissue from TA2 mice with SBC (×200, black arrowheads). d) Spontaneous intraductal carcinoma of TA2 mice (×100, black arrowheads). e) Spontaneous intraductal carcinoma of TA2 mice, the black arrow points at the intact basement membrane (×200, black arrowheads). f) Spontaneous invasive breast carcinoma of TA2 mice (×100, black arrowheads). g) The metastatic foci of lung in TA2 mice with SBC (×200, black arrowheads). h) The metastatic foci of liver in TA2 mice with SBC (×100, black arrowheads). i) This electron micrograph of SBC from TA2 mice showing the virus particles (×3000, black arrowheads). B. Expression of Wnt/β-catenin signaling pathway proteins. a) β-catenin expression in normal mammary tissue and the membranes of mammary gland epithelium were positive for β-catenin IHC staining (IHC, ×200, black arrowheads). b) β-catenin expression in precancerous lesion in TA2 mice and the cytoplasm of mammary gland epithelium were positive for β-catenin IHC staining (IHC, ×200). c) β-catenin expression of SBC tissue in TA2 mice and the nuclei of mammary gland epithelium were positive for β-catenin IHC staining (IHC, ×200). d) Wnt 5a expression was located in the cytoplasm of SBC in TA2 mice (IHC, ×200). e) GSK-3β expression was located in the cytoplasm of SBC in TA2 mice (IHC, ×200). f) Cyclin D1 expression was located in the nuclei of SBC in TA2 mice (IHC, ×200).

Article Snippet: After the nonspecific protein binding and the endogenous peroxidase activity were blocked, the sections were incubated with a rabbit polyclonal anti-β-catenin (Abcam, USA, dilution: 1:600), Wnt 5a (anti-rabbit, Beijing Bioscience Biotechnology Co. Ltd. Beijing, China; 1:100 dilution), GSK-3β (anti-rabbit, Beijing Bioscience Biotechnology Co. Ltd. Beijing, China; 1:100 dilution), and Cyclin D1 (ZA-0101, anti-rabbit, Beijing Zhongshan Biotechnology Co. Ltd. Beijing, China; 1:100 dilution) overnight at 4 °C.

Techniques: Membrane, Virus, Expressing, Immunohistochemistry

Journal: Pharmaceutical biology

Article Title: Yiguanjian decoction inhibits macrophage M1 polarization and attenuates hepatic fibrosis induced by CCl 4 /2-AAF.

doi: 10.1080/13880209.2021.1961820

Figure Lengend Snippet: Figure 3. YGJ inhibits the activation of M1 macrophages and non-canonical Wnt signalling pathways and inhibits the differentiation of hepatic progenitor cells into myofibroblasts in vivo. (a) STAT1, IRF3, IRF5, IRF8, and SOCS3 protein bands were depicted in the immunoblot images, and (b) the densitometric quantification of the protein bands presented as a histogram (n ¼ 5 per group). (c) The mRNA expressions of Wnt-4, -5 A, -5B, FZD-2, -3, and -6 in liver were measured by RT-PCR and nor- malized to GAPDH mRNA (n ¼ 5 per group). (d) Wnt5A, Wnt5B, and FZD2 protein bands were depicted in the immunoblot images, and (e) the densitometric quantifi- cation of the protein bands presented as a histogram (n ¼ 5 per group). (f) OV6, SOX9, EpCAM, CK19, and Hep mRNA expressions were measured by RT-PCR and normalized to GAPDH mRNA (n ¼ 5 per group). (g) EpCAM protein bands were depicted in the immunoblot images, and (h) the densitometric quantification of the protein bands presented as a histogram (n ¼ 5 per group). p < 0.05 and p < 0.01. N: untreated group (control); 2-AAF/CCl4: 2-acetylaminofluorene/carbon tetra- chloride-treated group; YGJ: 2-AAF/CCl4 þ Yiguanjian decoction-treated group; SORA: 2-AAF/CCl4 þ sorafenib-treated group.

Article Snippet: Rabbit polyclonal antibodies against Wnt5A and Wnt5B were purchased from Signalway Antibody LLC (MD, USA).

Techniques: Activation Assay, In Vivo, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control