Journal: European Journal of Medicinal Chemistry Reports

Article Title: Synthesis of new para-aminobenzoic acid derivatives, in vitro biological evaluation and preclinical validation of DAB-2-28 as a therapeutic option for the treatment of bladder cancer

doi: 10.1016/j.ejmcr.2022.100069

Figure Lengend Snippet: Fig. 3. Representative images (a) and graphical analysis (b) showing the immunodetection of phos- phorylated STAT3 (p-STAT3) in MB49-I cells pre- treated for 30 min with vehicle (DMSO) or DAB-1, DAB-2-28, DAB-2-31A and DAB-2-31B molecules at 15 and 30 μM, and then washed and recovered after 15 min of activation with 100 ng/mL IL6. The ratio of phosphorylated/unphosphorylated proteins was calculated from densitometric analysis of each sample to evaluate the relative activation of p-STAT3. *p < 0.05 and **p < 0.01 denote significant differences compared with vehicle control group.

Article Snippet: The antibodies against pSTAT3 (pY705; #9145), STAT3 (#4904), p-IκB (# 9246), iNOS (# 2977), and COX-2 (#12282) were purchased from Cell Signaling Technology (Danvers, MA) and β-actin (# A3854) was from Sigma Chemical Company (Oakville, Canada).

Techniques: Immunodetection, Activation Assay, Control

Journal: European Journal of Medicinal Chemistry Reports

Article Title: Synthesis of new para-aminobenzoic acid derivatives, in vitro biological evaluation and preclinical validation of DAB-2-28 as a therapeutic option for the treatment of bladder cancer

doi: 10.1016/j.ejmcr.2022.100069

Figure Lengend Snippet: Fig. 5. Representative images (a) and graphical analysis (b) showing the immunodetection of phos- phorylated STAT3 (p-STAT3) in MB49-I cells pre- treated for 30 min with vehicle (DMSO) or DAB-1, DAB-3-27, and DAB-3-33 molecules at 10, 20 and 30 μM, and then washed and recovered after 15 min of activation with 100 ng/mL IL6. The ratio of p-STAT3/ STAT3 proteins was calculated from densitometric analysis of each sample to evaluate the relative acti- vation of p-STAT3. *p < 0.05 and **p < 0.01 denote significant differences compared with vehicle control group.

Article Snippet: The antibodies against pSTAT3 (pY705; #9145), STAT3 (#4904), p-IκB (# 9246), iNOS (# 2977), and COX-2 (#12282) were purchased from Cell Signaling Technology (Danvers, MA) and β-actin (# A3854) was from Sigma Chemical Company (Oakville, Canada).

Techniques: Immunodetection, Activation Assay, Control

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Expression of Signal Transducer and Activator of Transcription 3 and Suppressor of Cytokine Signaling 3 in Urothelial Carcinoma

doi: 10.1016/s1607-551x(09)70569-8

Figure Lengend Snippet: Figure 1. Immunohistochemical staining of phosphorylated signal transducer and activator of transcription 3 (p-STAT3) Tyr705 and cytokine signaling 3 (SOCS3) in urothelial carcinomas. Nuclear staining of p-STAT3 (Tyr705): (A) low intensity grade (insert in A is normal urothelium); (B) high intensity grade. Cytoplasmic staining of SOCS3: (C) low intensity grade; (D) high intensity grade. Original magnification, 400×.

Article Snippet: The membranes were blocked with 5% non-fat dried milk in Tris-buffered Saline (pH 7.4) with Tween-20, and then incubated with either anti-phospho-STAT3 (Tyr705) (1:1,000 dilution; Cell Signaling Technology) or SOCS3 rabbit polyclonal primary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Immunohistochemical staining, Staining

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Expression of Signal Transducer and Activator of Transcription 3 and Suppressor of Cytokine Signaling 3 in Urothelial Carcinoma

doi: 10.1016/s1607-551x(09)70569-8

Figure Lengend Snippet: Figure 2. Western blotting of phosphorylated signal transducer and activator of transcription 3 (p-STAT3) Tyr705 and cytokine signaling 3 (SOCS3) protein in low-grade and high-grade UCs. p-STAT3 (Tyr705) expression was low and high in low- and high-grade UCs, respectively. Lane 1 = low-grade UC; lane 2=high-grade UC. However, the expression of SOCS3 was simi- lar in low- and high-grade UCs. GADPH protein expression was used as an internal control.

Article Snippet: The membranes were blocked with 5% non-fat dried milk in Tris-buffered Saline (pH 7.4) with Tween-20, and then incubated with either anti-phospho-STAT3 (Tyr705) (1:1,000 dilution; Cell Signaling Technology) or SOCS3 rabbit polyclonal primary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Western Blot, Expressing, Control

Journal: Brain research

Article Title: STAT3 protects dopaminergic neurons against degeneration in animal model of Parkinson's disease.

doi: 10.1016/j.brainres.2023.148691

Figure Lengend Snippet: Fig. 1. STAT3 reduced 6-OHDA neurotoxicity in N27 cell lines. Control N27 cells were examined in the absence (A) or presence (B) of 50 µM 6-OHDA for 24 h. N27 cells expressing caSTAT3 were examined in the absence (C) or presence (D) of 50 µM 6-OHDA for 24 h E) Graphical plot illustrating that caSTAT3 transfected N27 cells reduced percentage of cell deaths under 6-OHDA induced neurotoxicity. Three different batches of cell culture used. Result in mean ± SE. For each sample a minimum of 10,000 cells were recorded. The pink box represents the FVD + population of dead cells. The cells were double stained with FVD eFluor 660 and Annexin V-PE. The FVD-/Annexin V- population is regarded as normal healthy cells, while FVD-/AnnexinV + population is early apoptotic cells, and FVD+/AnnexinV+/- population represents necrotic/late apoptotic like cell death. Expression of caSTAT3 greatly reduced the percentage of dead cells 24 h after 6-OHDA treatment. F) Western blot analysis showed expression of phosphorylated STAT3 (pSTAT3) at Tyr705 (lane 3). An upregulation of pSTAT3 expression was found when compared to non-infected (lane 1) or GFP (lane 2) transfected N27 cell lysates. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Recombinant adeno-associated virus 2 (AAV2) carrying GFP or caRheb (kindly provided by Zhigang He, Harvard University, Boston, USA) was tagged with HA or caSTAT3 (pMXs-Stat3-C was a gift from Shinya Yamanaka (Addgene plasmid #13373) by helper virus-free system (Ayuso et al., 2010).

Techniques: Control, Expressing, Transfection, Cell Culture, Staining, Western Blot, Infection

Journal: Brain research

Article Title: STAT3 protects dopaminergic neurons against degeneration in animal model of Parkinson's disease.

doi: 10.1016/j.brainres.2023.148691

Figure Lengend Snippet: Fig. 2. caSTAT3 attenuated 6-OHDA-induced mitochondrial dysfunction. Control fluorescence, caSTAT3-expressing, and caRheb-expressing N27 cells were plated on coverslips and treated with indicated concentrations of 6-OHDA (µM) for 24 hrs prior to imaging (A); cells were loaded with 0.05 nM MitoSOX Red. B) The mean fluorescence intensity (F580 nm) of multiple cells (6 – 50 per condition) was quantified and plotted ± S.E.M. Two-way analysis of variance (ANOVA, F (2, 14) = 9.646, p = 0.0023) indicates significantly decreased mitochondrial superoxide production in caRheb or caSTAT3-expressing cells treated with either 6-OHDA dose compared to control. Sidak post hoc values for individual data points ** p < 0.01, *** p < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Recombinant adeno-associated virus 2 (AAV2) carrying GFP or caRheb (kindly provided by Zhigang He, Harvard University, Boston, USA) was tagged with HA or caSTAT3 (pMXs-Stat3-C was a gift from Shinya Yamanaka (Addgene plasmid #13373) by helper virus-free system (Ayuso et al., 2010).

Techniques: Control, Fluorescence, Expressing, Imaging

Journal: Brain research

Article Title: STAT3 protects dopaminergic neurons against degeneration in animal model of Parkinson's disease.

doi: 10.1016/j.brainres.2023.148691

Figure Lengend Snippet: Fig. 4. caSTAT3 protects dopaminergic neurons from 6-OHDA induced neurodegeneration. Data shows dopaminergic neuron survival 4 weeks after unilateral in jections of 6-OHDA in rats pre-injected with AAV/GFP (A,D), AAV/caSTAT3 (B,E,G) and AAV/caRheb (C,F) into the substantia nigra (SN). Coronal brain section (A,B, C) showing the extent of dopaminergic axon terminals with the striatum and the survival of dopaminergic neurons within the substantia nigra (D, E, F). Dramatic protection of striatal terminals and neurons is apparent in animals treated with either caSTAT3 (compare B & E to A & D, respectively) or caRheb (compare C & F to A & D, respectively). High magnification of TH + neurons from panel E shows neurons with a normal morphology (G). Quantitative analysis shows caRheb + caSTAT3, caSTAT3, or caRheb to statistically [ANOVA, F(3, 55) = 7.93, bonferroni post-hoc, p < 0.0001] preserve high density of dopaminergic nerve terminals within the striatum when compared to controls (H). Likewise, caRheb + caSTAT3, caSTAT3, or caRheb resulted in significantly increased survival [ANOVA, F(3, 81) = 21.16 bonferroni post-hoc, p < 0.0001] of dopaminergic neurons within the substantia nigra (I). Scale bar: A-F 500 µm; G 50 µm.

Article Snippet: Recombinant adeno-associated virus 2 (AAV2) carrying GFP or caRheb (kindly provided by Zhigang He, Harvard University, Boston, USA) was tagged with HA or caSTAT3 (pMXs-Stat3-C was a gift from Shinya Yamanaka (Addgene plasmid #13373) by helper virus-free system (Ayuso et al., 2010).

Techniques: Injection

Journal: Brain research

Article Title: STAT3 protects dopaminergic neurons against degeneration in animal model of Parkinson's disease.

doi: 10.1016/j.brainres.2023.148691

Figure Lengend Snippet: Fig. 5. Neuroprotection was observed in caSTAT3 transfected nigral dopami nergic neurons after 6-OHDA insult: A) Neurons are labelled with TH. B) Neurons are labelled with cMyc and C) neurons are merged for TH and cMyc. Data shows dopaminergic neuron survival 4 weeks after unilateral injections of 6-OHDA in rats pre-injected with AAV/caSTAT3. Scale bar = 50 µm.

Article Snippet: Recombinant adeno-associated virus 2 (AAV2) carrying GFP or caRheb (kindly provided by Zhigang He, Harvard University, Boston, USA) was tagged with HA or caSTAT3 (pMXs-Stat3-C was a gift from Shinya Yamanaka (Addgene plasmid #13373) by helper virus-free system (Ayuso et al., 2010).

Techniques: Transfection, Injection

Journal: Brain research

Article Title: STAT3 protects dopaminergic neurons against degeneration in animal model of Parkinson's disease.

doi: 10.1016/j.brainres.2023.148691

Figure Lengend Snippet: Fig. 6. caSTAT3 protects against 6-OHDA motor behavioral impairment: A) Amphetamine-induced rotational behavior was significantly decreased in caRheb + STAT3, caSTAT3, and caRheb expressing rats 4 weeks after 6-OHDA lesioning [ANOVA, F(3, 23) = 9.51, p = 0.0003] when compared to controls. B) After unilateral 6-OHDA lesions GFP treated animals showed a deficit in the use of the right (contralateral) paw, however, those treated with caRheb + caSTAT3, caSTAT3, or caRheb showed significant [ANOVA, F(3,17) = 8.29, p = 0.0013] use of the right paw in glass cylinder searching when compared to controls. Bonferroni post-hoc values for individual data points * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant adeno-associated virus 2 (AAV2) carrying GFP or caRheb (kindly provided by Zhigang He, Harvard University, Boston, USA) was tagged with HA or caSTAT3 (pMXs-Stat3-C was a gift from Shinya Yamanaka (Addgene plasmid #13373) by helper virus-free system (Ayuso et al., 2010).

Techniques: Expressing