Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: (A) Diagram of experimental design, representative FACS plots and quantification of CD9+CD26+ dermal fibroblasts percentage from Lin- Thy1+SCA1+ dorsal skin cells after three daily CNO intradermal injections from TRPV1 activated mice and their controls (****p<0.0001). (B) UMAP projection demonstrating the clustering of sorted Thy1+SCA1+ fibroblasts based on scRNAseq data, and the overlay of the Hashtag signal differentiating the subcutis and dermal clusters. (C) Normalized fold change (FC) calculation comparing cell numbers in dermal clusters between TRPV1 activated mice and their controls. (D) UMAP projection of sorted fibroblast scRNAseq data demonstrating cell distribution between clusters in the TRPV1 activated sample and control. (E-H) UMAP projection of sorted fibroblasts demonstrating the expression of (E) Col23A1 (F) Prdm1 (G) Crabp1 and (H) Lef1 as well as the list of top five velocity driver genes for cluster #3. (I) UMAP projection demonstrating calculated DP-core enrichment score and (J) Lepr expression. (K) UMAP projection of scRNASeq from Li et al. demonstrating its calculated DP-score (left) and cluster #3 enrichment score (right). (L) UMAP projection of sorted fibroblasts demonstrating the expression of Spp1. (M) Representative FACS histograms of anti-Spp1 staining of Lin- Thy1+SCA-1+ dermal cells (red) or CD9+CD26+ subset (orange) from the back skin of TRPV1 activated mouse. Secondary only staining control in blue. (N) Quantification of Spp1+CD9+CD26+ dermal fibroblasts from TRPV1 activated mice and controls (*p=0.017). (O) Representative images of RNAscope staining for Spp1 (green) and the Lepr (red) on dorsal skin collected from TRPV1 activated mice and controls. Scale bar, 100um. White arrows point out DP; yellow arrows point out Spp1 expression out of DP. DP images scale bar, 25um.

Article Snippet: InVivoMAb Anti-Spp1 neutralizing antibody (100D3) , BioXCell , Cat # #BE0372; RRID: AB_2927509.

Techniques: Control, Expressing, Staining, RNAscope

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: (A) Diagram of experimental design and representative images from shaved TRPV1 activated mice and controls. Mice monitored for hair coat growth for 12 days after the last TRPV1 activation with CNO (postnatal day 63). (B) Enlarged dashed rectangle from panel A depicting hair growth in a TRPV1 activated mouse. (C) HF length quantification of dorsal skin from mice treated as in A. Data combined from 6 to 8 HF per mouse, taken from 6 mice per group (****p<0.0001). (D) Representative immunofluorescent images of dorsal skin from mice treated as in A. Staining with anti-keratin14 (KRT14; green) marking epidermal layer and Ki67 (purple) marking the HF bulb. DAPI stain in blue. Scale bar 100μm. (E) Quantification of normalized Ki67 MFI signal in the peri-follicular area of dorsal skin from mice treated as in A. Data combined from 4 regions of interest (ROI) per mouse taken from 5 TRPV1 activated mice and 9 controls (****p<0.0001). (F) Diagram of experimental design and representative images from TRPV1 activated mice and controls treated with CNO on half of their back skin (marked with dashed rectangle). Anti-Spp1 neutralizing antibody or IgG control were injected intradermally to the same skin area. Data are combined from 4 HFs per mouse collected from 6 controls, 3 TRPV1 activated + IgG treated and 4 TRPV1 activated + anti-Spp1 treated mice (****p<0.0001). (G-H) Percentage of CD9+CD26+ dermal fibroblasts from TRPV1 activated mice and their controls intradermally injected with (G) QWF (SubP antagonist) or vehicle (**p=0.005, ***p=0.0004), (H) CGRP8–37 (CGRP antagonist) or vehicle (***p=0.0001, ****p<0.0001). (I) Diagram of experimental design and representative images from TRPV1 activated mice and controls pre-treated intradermally with CGRP8–37 or vehicle and monitored for hair coat growth for 14 days. Quantification of HF length from 6 to 8 HFs per mouse, taken from 3–6 mice per group (****p<0.0001). (J) Representative immunofluorescent images of dorsal skin from mice treated as in I. Stained with anti-KRT14 (green), Ki67 (pink), and DAPI (blue). Scale bar, 100μm.

Article Snippet: InVivoMAb Anti-Spp1 neutralizing antibody (100D3) , BioXCell , Cat # #BE0372; RRID: AB_2927509.

Techniques: Activation Assay, Staining, Control, Injection

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: (A) Representative immunofluorescent images and quantification of DRGs collected from mice with naïve back skin or 90 minutes after tape stripping. Stained with anti-c-Fos (red), anti-TRPV1 (green) and DAPI (blue). White arrowheads indicate neuron cell bodies that are TRPV1+ only. Data combined from 6–7 fields of view taken from 4 mice per group (****p<0.0001). Scale bar 50μm. (B) Percentage of CD9+CD26+ dermal fibroblasts from naïve back skin and 18 hours after tape stripping 6 or 12 times (***p=0.0004,****p<0.0001). (C) Diagram of experimental design and quantification of CD9+CD26+ dermal cell percentage 18 hours after tape stripping from the back skin of TRPV1Cre DTR+ treated with DTx or TRPV1Cre DTA+ mice and their controls (****p<0.0001). (D) Representative FACS histogram of anti-Spp1 staining. Dorsal skin treated as in C and gated on total Lin- Thy1+SCA-1+ cells (red) and the CD9+CD26+ cell subset (orange). Secondary only staining control in blue. (E) Quantification of Spp1+CD9+CD26+ dermal fibroblasts from TRPV1 ablated and control mice treated as in C (**p=0.005). (F) TRPV1 ablated mice and controls treated as in C and monitored for hair coat recovery for 12 days (postnatal day 61). (G) Representative bright field images of samples described in F. Data for HF length quantification collected from 8 HFs per mouse taken from 6 control and 5 TRPV1 ablated mice (****p<0.0001). (H) Ramp1 KO mice and controls treated as in F. HF length quantification from 4 HFs per mouse taken from 4 control and 5 Ramp1 KO mice (****p<0.0001). (I) Mice treated as in F and intradermally injected with anti-Spp1 or isotype control while being monitored for hair coat recovery for 16 days (postnatal day 65). HF length quantification from 4 HFs per mouse taken from 6 isotype control and 5 anti-Spp1 injected mice (**p=0.005).

Article Snippet: InVivoMAb Anti-Spp1 neutralizing antibody (100D3) , BioXCell , Cat # #BE0372; RRID: AB_2927509.

Techniques: Stripping Membranes, Staining, Control, Injection

Journal: Developmental cell

Article Title: Dermal TRPV1 innervations engage a macrophage and fibroblast containing pathway to activate hair growth in mice

doi: 10.1016/j.devcel.2024.05.019

Figure Lengend Snippet: Key resources table

Article Snippet: InVivoMAb Anti-Spp1 neutralizing antibody (100D3) , BioXCell , Cat # #BE0372; RRID: AB_2927509.

Techniques: Recombinant, RNAscope, Multiplex Assay, In Situ, Software

Journal: Cells

Article Title: Involvement of Cyclooxygenase-2 in Establishing an Immunosuppressive Microenvironment in Tumorspheres Derived from TMZ-Resistant Glioblastoma Cell Lines and Primary Cultures

doi: 10.3390/cells13030258

Figure Lengend Snippet: List of primary antibodies used in the present study.

Article Snippet: rabbit monoclonal anti-osteopontin , 1:1000 , Boster Biological Technology, Pleasanton, CA, USA.

Techniques:

Journal: Arthritis Research & Therapy

Article Title: Traditional Chinese medicine formula Bi-Qi capsule alleviates rheumatoid arthritis-induced inflammation, synovial hyperplasia, and cartilage destruction in rats

doi: 10.1186/s13075-018-1547-6

Figure Lengend Snippet: Collagen-induced arthritis (CIA) enhanced osteopontin (OPN) and cartilage oligomeric matrix protein (COMP) expression in paw joint cartilage, while the Bi-Qi capsule or methotrexate (MTX) treatment reduced this effect. Representative immunohistochemistry images of paw joint synovium with OPN and COMP immunostaining

Article Snippet: After blocking of non-specific binding sites with 10% normal goat serum for 1 h, the sections were incubated overnight at 4 °C with mouse-anti-COMP antibody (Boiss antibodies company, Beijing, China) in PBS or mouse anti-OPN antibody (Boster Bioengineering Co.Ltd., Wuhan, China) in PBS.

Techniques: Expressing, Immunohistochemistry, Immunostaining

Journal: Arthritis Research & Therapy

Article Title: Traditional Chinese medicine formula Bi-Qi capsule alleviates rheumatoid arthritis-induced inflammation, synovial hyperplasia, and cartilage destruction in rats

doi: 10.1186/s13075-018-1547-6

Figure Lengend Snippet: Bi-Qi capsule (BQ) or methotrexate (MTX) reduced collagen-induced arthritis (CIA)-induced osteopontin (OPN) and cartilage oligomeric matrix protein (COMP) upregulation in serum, and mRNA and protein levels in paw joint tissue. a, b OPN and COMP levels in serum. c, d OPN and COMP mRNA expression in paw joint cartilage. e, f OPN and COMP protein quantification from immunohistochemistry images of paw joint synovium. Data are presented as mean ± SD from eight rats in each group. Significant effect of treatment compared to control: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 . Significant effect of treatment compared to arthritic group: #### P < 0.0001 . Significant effect of treatment compared to high-dose BQ (BQ-HD): $$ P < 0.01, $$$$ P < 0.0001 . Significant effect of treatment compared to MTX-group: & P < 0.05, &&&& P < 0.0001. BQ-MD, moderate-dose BQ

Article Snippet: After blocking of non-specific binding sites with 10% normal goat serum for 1 h, the sections were incubated overnight at 4 °C with mouse-anti-COMP antibody (Boiss antibodies company, Beijing, China) in PBS or mouse anti-OPN antibody (Boster Bioengineering Co.Ltd., Wuhan, China) in PBS.

Techniques: Expressing, Immunohistochemistry, Control