Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Identification of genes regulated by leukemia-inhibitory factor in the mouse uterus at the time of implantation.

doi: 10.1210/me.2004-0110

Figure Lengend Snippet: Fig. 1. Regulation of Uterine IGFBP3 mRNA Expression by LIF Recombinant LIF was injected into one horn and PBS into the other horn of LIF/ females at 84 h of pseudopregnancy (n 10). IGFBP3 mRNA expression levels were determined in total uterine RNA isolated 12 h later by real-time RT-PCR. Values for IGFBP3 are normalized to 18S rRNA for each horn and expressed in arbitrary units relative to the level of the same gene in a standard RNA produced from mouse uterus. LIF treatment (solid bars) produced increased IGFBP3 expression in nine of 10 animals compared with the paired PBS-treated uterine horn (open bars). The mean increase was 2.3-fold and was statistically significant (P 0.02, paired Mann-Whitney).

Article Snippet: Two ip injections each of 5 g recombinant human LIF (R&D systems 250-LF) in PBS were given at 0900 h and 1600 h on d 4 of pregnancy, and the animals were allowed to proceed to term.

Techniques: Expressing, Recombinant, Injection, Isolation, Quantitative RT-PCR, Produced, MANN-WHITNEY

Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Identification of genes regulated by leukemia-inhibitory factor in the mouse uterus at the time of implantation.

doi: 10.1210/me.2004-0110

Figure Lengend Snippet: Fig. 6. LIF Regulation of Amphiregulin and IRG1 in the Uterus Recombinant LIF was injected into one horn and PBS into the other horn of LIF/ females at 84 h of pseudopreg- nancy (n 10, same animals as shown in Fig. 1). Amphiregu- lin and IRG1 mRNA expression levels were determined in total uterine RNA isolated 12 h later by real-time RT-PCR. Values for each mRNA species are normalized to 18S rRNA for each horn and expressed in arbitrary units relative to the level of the same gene in a standard RNA produced from mouse uterus. LIF treatment (solid bars) increased both am- phiregulin and IRG1 expression in 10 of 10 animals compared with the corresponding PBS-treated horn (open bars). The mean increase for amphiregulin was 7.3-fold and for IRG1 was 3.7-fold and was statistically significant for either gene (P 0.002, paired Mann-Whitney test).

Article Snippet: Two ip injections each of 5 g recombinant human LIF (R&D systems 250-LF) in PBS were given at 0900 h and 1600 h on d 4 of pregnancy, and the animals were allowed to proceed to term.

Techniques: Recombinant, Injection, Expressing, Isolation, Quantitative RT-PCR, Produced, MANN-WHITNEY

Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

Article Title: TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

doi: 10.1111/wrr.70032

Figure Lengend Snippet: FIGURE 1 TWEAK/Fn14 signalling upregulates the expressions of HFSC markers. (A) Immunofluorescence analysis of Fn14 and CXCR4 expression in mouse hair follicles. The white arrows indicate the Fn14- or CXCR4-positive cells; however, these positive cells were significantly diminished and even became indistinct in the Fn14-deficient specimen. Scale bar = 20 μm. (B) Histochemical analysis of Fn14, K19 and CD34 expression in mouse hair follicles. Scale bar = 20 μm. (C) Primary human HFSCs cultured in vitro. (D) Immunofluorescence analysis of β1 integrin and K15 expression in HFSCs stimulated with TWEAK (10 ng/mL). Scale bar = 10 μm. (e, f) Western blotting of K19, integrin β1 and K15 proteins. Data represent mean ± SEM from three independent experiments. *p < 0.05, compared with the 0 ng/mL TWEAK group. CXCR4, chemokine (C X C motif) receptor 4; Fn14, fibroblast growth factor-inducible 14; HFSC, hair follicle stem cells; K15/K19, keratins 15/19; TWEAK, tumour necrosis factor- like weak inducer of apoptosis.

Article Snippet: HFSCs were stimulated for 48 hours with recombinant human TWEAK (0–250 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: Immunofluorescence, Expressing, Cell Culture, In Vitro, Western Blot

Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

Article Title: TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

doi: 10.1111/wrr.70032

Figure Lengend Snippet: FIGURE 2 TWEAK induces proliferation, migration and cytokine production in HFSCs. Human HFSCs were cultured in vitro and stimulated with TWEAK (0–250 ng/mL). (A) Proliferation of HFSCs analysed via flow cytometry. n = 5 per group. (B) Cell migration assessed via scratch analysis. n = 5 per group. (C) mRNA expression levels of EGF, BFGF, TGF-β, NGF and VEGF measured via qRT-PCR. n = 3 per group. (D) Cytokine levels in the supernatants determined via ELISA. n = 3 per group. Data represent mean ± SEM from three to five independent experiments. *p < 0.05 compared with the 0 ng/mL group. #p < 0.05 compared with the 10 ng/mL group. HFSC, hair follicle stem cells; qRT-PCR, quantitative real-time PCR; SEM, standard error of the mean; TWEAK, tumour necrosis factor-like weak inducer of apoptosis.

Article Snippet: HFSCs were stimulated for 48 hours with recombinant human TWEAK (0–250 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: Migration, Cell Culture, In Vitro, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

Article Title: TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

doi: 10.1111/wrr.70032

Figure Lengend Snippet: FIGURE 3 TWEAK upregulates the expressions of HFSC phenotype markers. Human HFSCs were cultured in vitro and stimulated with TWEAK (0–250 ng/mL). (A) Fn14 and TNFR2 expressions detected via immunofluorescence. (B) mRNA expression levels of FN14, TNFR2, IGFR and CXCR4 analysed via qRT-PCR. (C, D) Protein expressions of these markers determined via Western blotting, with quantification using ImageJ software. Data represent mean ± SEM from three independent experiments. *p < 0.05 compared with the 0 ng/mL group. #p < 0.05 compared with the 10 ng/mL group. Δp < 0.05 compared with the 50 ng/mL group. HFSC, hair follicle stem cells; qRT-PCR, quantitative real-time PCR; SEM, standard error of the mean; TWEAK, tumour necrosis factor-like weak inducer of apoptosis.

Article Snippet: HFSCs were stimulated for 48 hours with recombinant human TWEAK (0–250 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: Cell Culture, In Vitro, Immunofluorescence, Expressing, Quantitative RT-PCR, Western Blot, Software, Real-time Polymerase Chain Reaction

Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

Article Title: TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

doi: 10.1111/wrr.70032

Figure Lengend Snippet: FIGURE 4 TWEAK activates Wnt/β-catenin signalling in HFSCs. Human HFSCs were cultured in vitro and stimulated with TWEAK (10 ng/mL). (A) mRNA expression levels of WNT5A, CTNNB and GSK3B analysed via qRT-PCR. (B, C) Protein expressions of these markers detected via Western blotting, with quantification using ImageJ software. (D) mRNA expression levels of TNFR2, IGFR and CXCR4 analysed in cells pretreated with the β-catenin inhibitor XAV-939. (E, F) Protein expressions of these markers detected via Western blotting, with quantification using ImageJ software. Data represent mean ± SEM from three independent experiments. In (A, C), *p < 0.05 compared with the blank group. In (D, F), *p < 0.05 compared with the PBS alone group; #p < 0.05 compared with the XAV-939 group. Δp < 0.05 compared with the TWEAK alone group. HFSC, hair follicle stem cells; qRT-PCR, quantitative real-time PCR; TWEAK, tumour necrosis factor- like weak inducer of apoptosis.

Article Snippet: HFSCs were stimulated for 48 hours with recombinant human TWEAK (0–250 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: Cell Culture, In Vitro, Expressing, Quantitative RT-PCR, Western Blot, Software, Real-time Polymerase Chain Reaction

Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society

Article Title: TWEAK regulates the functions of hair follicle stem cells via the Fn14-Wnt/β-catenin-CXCR4 signalling axis.

doi: 10.1111/wrr.70032

Figure Lengend Snippet: FIGURE 5 CXCR4 inhibitor suppresses the effect of TWEAK on HFSCs. Human HFSCs were cultured in vitro and treated with TWEAK (10 ng/mL) in the presence or absence of the CXCR4 inhibitor EPI-X4. (A) Proliferation of HFSCs assessed via flow cytometry. n = 5 per group. (B, C) Protein expressions of differentiation markers detected via Western blotting and quantitated using ImageJ software. n = 3 per group. (D). Cytokines synthesised by HFSCs and measured via ELISA in the culture supernatants. n = 3 per group. Data represent mean ± SEM from three to five independent experiments. *p < 0.05 compared with the blank group. #p < 0.05 compared with the TWEAK alone group. CXCR4, chemokine (C X C motif) receptor 4; HFSC, hair follicle stem cells; TWEAK, tumour necrosis factor-like weak inducer of apoptosis.

Article Snippet: HFSCs were stimulated for 48 hours with recombinant human TWEAK (0–250 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: Cell Culture, In Vitro, Flow Cytometry, Western Blot, Software, Enzyme-linked Immunosorbent Assay