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: iScience

Article Title: BMAL1 upregulates STX17 levels to promote autophagosome-lysosome fusion in hippocampal neurons to ameliorate Alzheimer's disease

doi: 10.1016/j.isci.2024.111413

Figure Lengend Snippet: Figure 5. The AD model showed circadian rhythm disturbance and decreased Bmal1 expression (A) The protocol of synchronize in vivo and in vitro. (B) APP/PS1 mice displayed circadian rhythm disorders, increased daytime activity, and prolonged free-running cycles.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-LC3B antibody Abcam Cat #ab192890 BMAL1 (D2L7G) Rabbit mAb Cell Signaling Technology Cat #14020S STX17 Polyclonal antibody Proteintech Cat #17815-1-AP VAMP8 Polyclonal antibody Proteintech Cat #15546-1-AP SNAP29 Polyclonal antibody Proteintech Cat #12704-1-AP Anti-SQSTM1 / p62 antibody Abcam Cat #ab109012 Anti-Amyloid Precursor Protein antibody Abcam Cat #ab32136 Bacterial and virus strains Lentivirus-overexpressing STX17 Shanghai Genechem N/A Lentivirus-overexpressing BMAL1 Shanghai Genechem N/A STX17-siRNA Hanbio Biotechnology N/A Chemicals, peptides, and recombinant proteins RNAiso Plus TaKaRa Cat #9109 RIPA buffer Boster Cat #AR0102 Glutaraldehyde,2.5%(EM Grade) Solarbio Cat #P1126 Critical commercial assays PrimeScript RT Master Mix TaKaRa Cat #RR036A SYBR Premix Ex TaqTMII TaKaRa Cat #DRR041A Experimental models: Cell lines HT22 Chinese Academy of Sciences Cell Bank CSTR:19375.09.3101MOUGNM47 Experimental models: Organisms/strains SPF mice Model Animal Research Center APP/PS1 transgenic mice Software and algorithms ImageJ ImageJ https://imagej.nih.gov/ij/ GraphPad Prism GraphPad Prism https://www.graphpad.com/ Other Code of GSE21779 GEO https://www.ncbi.nlm.nih.gov/gds/ Code for data analysis and visualization bioinformatics https://www.bioinformatics.com.cn Code for revealed the binding sites JASPAR http://jaspar.genereg.net/ Data: Western Blot and Microscopy data Lead contact 163.wangxh@163.com

Techniques: Expressing, In Vivo, In Vitro, Activity Assay

Journal: iScience

Article Title: BMAL1 upregulates STX17 levels to promote autophagosome-lysosome fusion in hippocampal neurons to ameliorate Alzheimer's disease

doi: 10.1016/j.isci.2024.111413

Figure Lengend Snippet: Figure 6. BMAL1 regulates STX17 to affect autophagy and amyloid deposition (A) JASPAR analysis revealed the recognition sites of BMAL1 on the promoter sequence of STX17. (B) Detection of luciferase activity after the STX17 promoter sequence plasmid and BMAL1 plasmid were transfected into HT22 cells. (C) Autophagic flow was partially restored in APP-overexpressed HT22 after BMAL1 overexpression. (D) Amyloid deposition decreases after BMAL1 overexpression. n = 6; scale bar, 50 mm. *p < 0.05 vs. the Lv-OE-APP+Lv-NC group; **p < 0.01 vs. the Lv-OE- APP+Lv-NC group. Data are represented as mean ± SD.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-LC3B antibody Abcam Cat #ab192890 BMAL1 (D2L7G) Rabbit mAb Cell Signaling Technology Cat #14020S STX17 Polyclonal antibody Proteintech Cat #17815-1-AP VAMP8 Polyclonal antibody Proteintech Cat #15546-1-AP SNAP29 Polyclonal antibody Proteintech Cat #12704-1-AP Anti-SQSTM1 / p62 antibody Abcam Cat #ab109012 Anti-Amyloid Precursor Protein antibody Abcam Cat #ab32136 Bacterial and virus strains Lentivirus-overexpressing STX17 Shanghai Genechem N/A Lentivirus-overexpressing BMAL1 Shanghai Genechem N/A STX17-siRNA Hanbio Biotechnology N/A Chemicals, peptides, and recombinant proteins RNAiso Plus TaKaRa Cat #9109 RIPA buffer Boster Cat #AR0102 Glutaraldehyde,2.5%(EM Grade) Solarbio Cat #P1126 Critical commercial assays PrimeScript RT Master Mix TaKaRa Cat #RR036A SYBR Premix Ex TaqTMII TaKaRa Cat #DRR041A Experimental models: Cell lines HT22 Chinese Academy of Sciences Cell Bank CSTR:19375.09.3101MOUGNM47 Experimental models: Organisms/strains SPF mice Model Animal Research Center APP/PS1 transgenic mice Software and algorithms ImageJ ImageJ https://imagej.nih.gov/ij/ GraphPad Prism GraphPad Prism https://www.graphpad.com/ Other Code of GSE21779 GEO https://www.ncbi.nlm.nih.gov/gds/ Code for data analysis and visualization bioinformatics https://www.bioinformatics.com.cn Code for revealed the binding sites JASPAR http://jaspar.genereg.net/ Data: Western Blot and Microscopy data Lead contact 163.wangxh@163.com

Techniques: Sequencing, Luciferase, Activity Assay, Plasmid Preparation, Transfection, Over Expression