Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeted Delivery of CD34 Aptamer-Coupled Tocilizumab Microspheres for Effective Treatment of Thyroid-Associated Ophthalmopathy

doi: 10.1167/iovs.66.11.57

Figure Lengend Snippet: Tocilizumab/(anti-mmu-IL-6R) inhibits STAT3 pathway activation in fibroblasts and orbital tissues . ( A ) The protein levels of p-STAT3 and STAT3 levels in differentially treated human orbital fibroblasts were determined using immunoblotting. ( B ) The protein levels of STAT3 and p-STAT3 levels in model mice orbital tissues were determined using immunoblotting ( N = 3 biological replicates). * P < 0.05, ** P < 0.01.

Article Snippet: Primary antibodies against p-STAT3 (CSB-RA022812A727phHU, Cusabio), STAT3 (10253-2-AP, Proteintech), FGF2 (11234-1-AP, Proteintech), FGFR2 (CSB- PA000992 , Cusabio), and PCNA (10205-2-AP, Proteintech) were then incubated overnight.

Techniques: Activation Assay, Western Blot

Journal: Pharmaceuticals

Article Title: Multiple Antitumor Molecular Mechanisms Are Activated by a Fully Synthetic and Stabilized Pharmaceutical Product Delivering the Active Compound Sulforaphane (SFX-01) in Preclinical Model of Human Glioblastoma

doi: 10.3390/ph14111082

Figure Lengend Snippet: Immuno-histochemical and enzymatic evaluation on markers of proliferation, apoptosis, autophagy, angiogenesis, stemness and signaling in untreated (control) and treated with SFX-01 in U87MG and T98G xenografts.

Article Snippet: Antibodies against p21 (STJ94862), p27 (STJ94867), cyclin B1 (STJ97479), cyclin D1 (STJ92538), CDK4 (STJ11101180), CD44 (STJ119186), nestin (STJ113858), N-cadherin (STJ94353), catenin-beta (STJ92042), Bax (STJ118092), phospho-Akt (S473) (STJ90166), total Akt (STJ91538), 4E-BP1 (STJ91385), phospho-4E-BP1 (S65) (STJ90778), STAT3 (STJ95808), and phospho-STAT3 (Tyr 705 code STJ90411) were purchased from St John’s Laboratory Ltd. (Docklands Campus University Way, London, UK).

Techniques: Control, TUNEL Assay, Activity Assay

Journal: Cell reports

Article Title: Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration

doi: 10.1016/j.celrep.2014.08.047

Figure Lengend Snippet: (A) Immunofluorescence on retinal sections shows activated p-Stat3 expression in BrdU+ MG-derived progenitors that are localized to the injury site at 4 dpi. (B) Schematic of gfap:stat3-gfp transgene construct shows the fusion gene, stat3-gfp, under control of the gfap promoter regulatory elements. (C) In gfap:stat3-gfp transgenic fish, Stat3-GFP fusion protein expression is undetectable in MG of the uninjured eye, while restricted to MG-derived progenitors at the injury site at 4 dpi. (D) Confocal images showing co-localization of Stat3-GFP with glutamine synthetase (GS)+/p-Stat3+/BrdU+ MG-derived progenitors at 4 dpi. In (A and C) the asterisk marks the injury site (needle poke) and arrows point to MG-derived progenitors. In (C) white dots indicate autofluorescence unique to the green channel (see Figure S2G). Error bars, s.d. Scale bars, 50 µm (A, C) and 20 µm (D). INL, inner nuclear layer; ONL, outer nuclear layer; RPE, retinal pigment epithelium; dpi, days post injury. See also Figures S1and S2.

Article Snippet: Immunofluorescence and Western Blots Anti-GFP and 4C4 immunofluorescence were as previously described ( Craig et al., 2010 ; Fausett and Goldman, 2006 ). p-Stat3 immunoflourescence was performed using mouse anti-phospho-zebrafish Stat3 (Tyr708) antibody (MBL) at 1:100 dilution.

Techniques: Immunofluorescence, Expressing, Derivative Assay, Construct, Control, Transgenic Assay

Journal: Cell reports

Article Title: Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration

doi: 10.1016/j.celrep.2014.08.047

Figure Lengend Snippet: (A) GFP immunofluorescence in 6-ascl1a:gfp fish shows that Jak inhibitors, P6 and JSI-124, applied at the time of retinal injury, inhibit injury-dependent transgene induction. (B) Diagram of ascl1a promoter constructs used to generate transgenic lines. (C) GFP immunofluorescence shows that a distal 1.5 kb fragment of the ascl1a promoter is required for injury-dependent transgene expression and that both consensus Stat3 sites located in this promoter fragment are necessary for this expression. BrdU+ cells indicate the injury site and a normal regenerative response. The asterisk marks the injury site (needle poke). Scale bar, 50 µm. INL, inner nuclear layer; dpi, days post injury. See also Figure S4.

Article Snippet: Immunofluorescence and Western Blots Anti-GFP and 4C4 immunofluorescence were as previously described ( Craig et al., 2010 ; Fausett and Goldman, 2006 ). p-Stat3 immunoflourescence was performed using mouse anti-phospho-zebrafish Stat3 (Tyr708) antibody (MBL) at 1:100 dilution.

Techniques: Immunofluorescence, Construct, Transgenic Assay, Expressing

Journal: Cell reports

Article Title: Leptin and IL-6 Family Cytokines Synergize to Stimulate Müller Glia Reprogramming and Retina Regeneration

doi: 10.1016/j.celrep.2014.08.047

Figure Lengend Snippet: (A) Intravitreal injection of Leptin/CNTF, or Leptin/IL-11 into the eye of gfap:stat3-gfp fish stimulates Stat3-GFP expression and BrdU incorporation throughout the uninjured retina’s INL, while Leptin, CNTF or IL-11 alone had little effect. Note that GFP reports activated p-Stat3 expression (Figure 1; Figure S2). White dots indicate autofluorescence unique to the green channel (see Figure S7K). The arrows point to MG-derived progenitors in the top 4 panels. (B) Quantification of Stat3-GFP+ cells in (A); ***P<0.001 (combination vs individual), n=4. (C) Quantification of BrdU+ cells in (A); ***P<0.001 (combination vs individual), n=4. (D) Western blot shows that retinal injury or intravitreal injection of cytokines into uninjured eye increases p-Stat3 expression. GS serves as loading control. (E) A model showing Leptin and IL-6 family cytokines synergize to stimulate MG reprogramming and retina regeneration via a Jak/Stat3 signaling pathway, which is essential for activating reprogramming genes like hbegf and ascl1a. Error bars, s.d. Scale bars, 50 µm. ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer.

Article Snippet: Immunofluorescence and Western Blots Anti-GFP and 4C4 immunofluorescence were as previously described ( Craig et al., 2010 ; Fausett and Goldman, 2006 ). p-Stat3 immunoflourescence was performed using mouse anti-phospho-zebrafish Stat3 (Tyr708) antibody (MBL) at 1:100 dilution.

Techniques: Injection, Expressing, BrdU Incorporation Assay, Derivative Assay, Western Blot, Control

Journal: Journal of Neuroinflammation

Article Title: TRPV1 mediates astrocyte activation and interleukin-1β release induced by hypoxic ischemia (HI)

doi: 10.1186/s12974-019-1487-3

Figure Lengend Snippet: Antibodies for Western blot

Article Snippet: STAT3 , Rabbit , Hangzhou HuaAn Biotech , ET1607-38 , 1:1000 , Overnight 4 °C.

Techniques: Western Blot

Journal: Journal of Neuroinflammation

Article Title: TRPV1 mediates astrocyte activation and interleukin-1β release induced by hypoxic ischemia (HI)

doi: 10.1186/s12974-019-1487-3

Figure Lengend Snippet: Knocking out TRPV1 inhibited the phosphorylation of JAK2 and STAT3, astrocytes activation, and the expression of IL-1β. a Western blot analysis showed the upregulated after OGD, and knocking out TRPV1 or pretreating with capsaicin inhibited the phosphorylation of JAK2 and STAT3 compared to OGD group. b , c # P < 0.05 (Tukey’s test after two-way repeated-measures ANOVA). n = 5 for each group. d The graphical representation of the fold changes of the expression of GFAP, treating with Stattic (5 μM) inhibited the upregulation of GFAP. n = 5 for each group. e Western blot revealed treating with Stattic (5 μM) inhibited the upregulation of GFAP. n = 5 for each group. f As confirmed by quantification of independent blots. # P < 0.05 (Tukey’s test after two-way repeated-measures ANOVA). n = 5 for each group. g , h The graphical representation of the fold changes of IL-1β. Treating with Stattic (5 μM) inhibited the upregulation of IL-1β. n = 6 for each group. Cap, 10 μM capsaicin. Sta, 5 μM stattic. Average values represent the mean ± SEM. ** P < 0.01, *** P < 0.001 versus control (Tukey’s test after one-way ANOVA). # P < 0.05, ## P < 0.01 versus WT-OGD (Tukey’s test after one-way ANOVA). ns P > 0.05 (Tukey’s test after one-way ANOVA)

Article Snippet: STAT3 , Rabbit , Hangzhou HuaAn Biotech , ET1607-38 , 1:1000 , Overnight 4 °C.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Journal of Neuroinflammation

Article Title: TRPV1 mediates astrocyte activation and interleukin-1β release induced by hypoxic ischemia (HI)

doi: 10.1186/s12974-019-1487-3

Figure Lengend Snippet: Schematic illustration of the proposed mechanism. TRPV1 plays an important role in neonatal HI-induced astrocytes activation and the transcription and post-translational protein modification of IL-1β. (1) TRPV1 stimulated the phosphorylation of JAK2, and phosphorylated JAK2 promoted the phosphorylation of STAT3. And activated STAT3 bound to its target genes (GFAP and IL-1β) and upregulated their expression. (2) TRPV1 promoted the formation of NLRP3 inflammasome (NLRP3, ASC, and caspase-1), which mediated the activation of caspase-1 and then stimulated the cleavage of pro-IL-1β

Article Snippet: STAT3 , Rabbit , Hangzhou HuaAn Biotech , ET1607-38 , 1:1000 , Overnight 4 °C.

Techniques: Activation Assay, Modification, Expressing