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: The Journal of Neuroscience

Article Title: Neuroprotective and Axon Growth-Promoting Effects following Inflammatory Stimulation on Mature Retinal Ganglion Cells in Mice Depend on Ciliary Neurotrophic Factor and Leukemia Inhibitory Factor

doi: 10.1523/JNEUROSCI.2770-09.2009

Figure Lengend Snippet: Changes in the expression of CNTF and LIF, as well as JAK/STAT3 pathway activation, in the murine retina after ONC and LI. A, Immunohistochemical staining of an untreated control retina (con), a retina 5 d after optic nerve crush (onc), and onc + lens injury (li) from wild-type mice as indicated using an anti-CNTF (red) and an anti-GFAP (green) antibody. Scale bar, 50 μm. GCL, ganglion cell layer; INL, inner nuclear layer. B, Western blot analysis of retinal lysates from wild-type (wt) and CNTF-deficient (cntf−/−) mice 5 d after onc, onc + li, or no previous treatment (con) using specific antibodies against GFAP, GAP43, LIF, phospho-STAT3 (pSTAT3), or CNTF. Tubulin served as a loading control. C, Immunohistochemical staining of astrocytes of an untreated control retina, a retina 5 d after onc, and after onc + li of wt and cntf−/− mice using an anti-LIF (R&D) (red) and anti-GFAP antibody (green). Scale bar, 25 μm. D, Quantitative real-time PCR. LIF expression levels were quantified relative to GAPDH in wt and cntf−/− mice 5 d after onc, onc + li, or untreated animals (−). *p < 0.05; ***p < 0.001. E, Western blot analysis of retinal lysates from CNTF/LIF double knock-out (cntf−/−/lif−/−) mice 5 d after onc, onc + li, or no previous treatment (−) using specific antibodies against GAP43, GFAP, and pSTAT3. Tubulin expression verified that the same amount of retinal protein was loaded per lane.

Article Snippet: C , Immunohistochemical staining of astrocytes of an untreated control retina, a retina 5 d after onc, and after onc + li of wt and cntf −/− mice using an anti-LIF (R&D) (red) and anti-GFAP antibody (green).

Techniques: Expressing, Activation Assay, Immunohistochemical staining, Staining, Western Blot, Real-time Polymerase Chain Reaction, Knock-Out

Journal: The Journal of Neuroscience

Article Title: Neuroprotective and Axon Growth-Promoting Effects following Inflammatory Stimulation on Mature Retinal Ganglion Cells in Mice Depend on Ciliary Neurotrophic Factor and Leukemia Inhibitory Factor

doi: 10.1523/JNEUROSCI.2770-09.2009

Figure Lengend Snippet: Regenerative state of RGCs 5 d after ONC and ONC + LI in mature wild-type and CNTF-deficient mice. A, Dissociated retinal cell cultures immunostained with an antibody against βIII-tubulin, showing spontaneously regenerating RGCs after 24 h in culture. Wild-type (wt) and CNTF-deficient (cntf−/−) mice had received ONC (onc), ONC + LI (onc + li), or no treatment (con) 5 d previously. Scale bar, 50 μm. B, Quantitation of neurite outgrowth of groups in A and of CNTF/LIF double knock-out (cntf−/−/lif−/−) mice 5 d after ONC + LI, indicated as average neurite length per RGC. C, Quantitation of RGCs per well after 24 h in culture, demonstrating no significant differences between groups. Significance between groups: ***p < 0.001; ns., nonsignificant.

Article Snippet: C , Immunohistochemical staining of astrocytes of an untreated control retina, a retina 5 d after onc, and after onc + li of wt and cntf −/− mice using an anti-LIF (R&D) (red) and anti-GFAP antibody (green).

Techniques: Quantitation Assay, Knock-Out

Journal: The Journal of Neuroscience

Article Title: Neuroprotective and Axon Growth-Promoting Effects following Inflammatory Stimulation on Mature Retinal Ganglion Cells in Mice Depend on Ciliary Neurotrophic Factor and Leukemia Inhibitory Factor

doi: 10.1523/JNEUROSCI.2770-09.2009

Figure Lengend Snippet: Axon regeneration and survival of RGCs in vivo after LI in wild-type, CNTF-deficient and CNTF/LIF double knock-out mice. A, Longitudinal sections through the optic nerve showing GAP43-positive axons distal to the injury site (asterisk) in wild-type (wt), CNTF-deficient (cntf −/−) and CNTF/LIF double knock-out (cntf −/−/lif −/−) mice 2 weeks after ONC alone (onc) or ONC + LI (onc + li). Scale bar, 100 μm. B, Quantification of axon regeneration (number of axons growing 0.25, 0.5, and 1 mm beyond the injury site per optic nerve) 2 weeks after ONC alone or ONC + LI. C, Quantification of surviving RGCs (βIII-tubulin-positive RGCs per retinal cross-section) 2 weeks after ONC alone, ONC + LI, or untreated retinas (−). *p < 0.05; *** p < 0.001.

Article Snippet: C , Immunohistochemical staining of astrocytes of an untreated control retina, a retina 5 d after onc, and after onc + li of wt and cntf −/− mice using an anti-LIF (R&D) (red) and anti-GFAP antibody (green).

Techniques: In Vivo, Knock-Out

Journal: The Journal of Neuroscience

Article Title: Neuroprotective and Axon Growth-Promoting Effects following Inflammatory Stimulation on Mature Retinal Ganglion Cells in Mice Depend on Ciliary Neurotrophic Factor and Leukemia Inhibitory Factor

doi: 10.1523/JNEUROSCI.2770-09.2009

Figure Lengend Snippet: Neurite growth-promoting effects of LIF and CNTF on mature RGCs in culture. A, Quantitation of neurite outgrowth of RGCs in the presence of increasing concentrations of LIF (as indicated), CNTF and an anti-LIF-antibody (α-LIF) for 3 d. Values were normalized to untreated controls. Treatment effects compared with the group treated with vehicle only. *p < 0.05; ***p < 0.001. B, Quantitation of RGCs per well after 3 d in culture, demonstrating no significant differences between groups.

Article Snippet: C , Immunohistochemical staining of astrocytes of an untreated control retina, a retina 5 d after onc, and after onc + li of wt and cntf −/− mice using an anti-LIF (R&D) (red) and anti-GFAP antibody (green).

Techniques: Quantitation Assay