Journal: ACS Omega

Article Title: Site-Specific Glycan Conjugation Improves Stability and Efficacy of an Antibody–Drug Conjugates Bearing DXd as a Cytotoxic Payload

doi: 10.1021/acsomega.5c10898

Figure Lengend Snippet: Preparation Scheme of Glycan-Conjugated ADC and Payload Structures. (i) Inherent Heterogeneous Glycan Removal by ENGase. ENGase Cleaves the Bond Between the β-1,4-Glycosidic Linkage Within the N,N ′-Diacetylchitobiose Core. Heterogeneous Regions are Indicated in Parentheses; (ii) An Azide-Carrying Glycan 4 was Added via the ENGase Mutant; (iii) Biorthogonal Reaction Between the Azide Group and DBCO. Payload Structure: DBCO-PEG 3 -VC-PAB-MMAE 5 ; DBCO-PEG 4 -GGFG-DXd 6 ; Trasuzumab-SG-PEG 3 -VC-PAB-MMAE 7 ; Trasuzumab-SG-PEG 4 -GGFG-DXd 8

Article Snippet: DBCO-PEG 3 -VC-PAB-MMAE 5 and DBCO-PEG 4 -Gly-Gly-Phe-Gly-DXd 6 were purchased from MedChemExpress (Monmouth Junction, NJ, USA) and Levena Biopharma (San Diego, CA, USA), respectively.

Techniques: Glycoproteomics, Mutagenesis

Journal: European journal of pharmacology

Article Title: Terazosin attenuates abdominal aortic aneurysm formation by downregulating Peg3 expression to inhibit vascular smooth muscle cell apoptosis and senescence.

doi: 10.1016/j.ejphar.2024.176397

Figure Lengend Snippet: Fig. 3. TZ reduces AAA formation by attenuating PEG3 signaling. (A) Heatmap depicting differentially expressed genes between control and 10 nM TZ-treated MOVAS cells. (B) Representative images of IHC staining for PEG3 in human AAA samples (scale bar, 50 μm) and quantification of IHC results (n = 3). Black arrows indicate PEG3-positive cells. *p < 0.05 compared with the control group. (C) MOVAS cells were treated by 1 μM Ang II with or without TZ for 48 h. Representative western blots and quantification of PEG3 and MMP2 protein expressions in MOVAS (n = 3–6). *p < 0.05 compared with ctrl group, #p < 0.05 between Ang-TZ and Ang II group. (D) (Upper) Representative IHC staining images of PEG3 in Apoe−/−mice suprarenal aortas from each group (scale bar, 50 μm) and quantification of IHC results (n = 5–9). *p < 0.05 compared with the sham-water group, #p < 0.05 compared with the Ang II-water group. (Lower) Representative IHC staining images of PEG3 in C57BL/6J mice infrarenal aortas from each group (scale bar, 50 μm) and quantification of IHC results (n = 5–12). *p < 0.05 compared with the sham-water group, #p < 0.05 compared with the CaCl2-water group. Black arrows indicate PEG3-positive cells. (E) (Left) Representative Immunofluorescent staining images of PEG3 (Green), α-SMA (Red) and DAPI (Blue) in Apoe−/−mice suprarenal aortas from sham-water group, Ang II-water group elastin unbroken area, Ang II- water group elastin broken area and Ang II-TZ (100 μg//kg) group (scale bar, 50 μm). (Right) Representative Immunofluorescent staining images of PEG3 (Green), α-SMA (Red) and DAPI (Blue) in C57BL/6J mice infrarenal aortas from the indicated groups (scale bar, 50 μm). IHC, Immunohistochemistry. α-SMA, α-smooth muscle actin.

Article Snippet: Primary antibodies used included: PEG3 (AF9152, 1:150, Affinity, China), α-SMA (BM0002, 1:200, BOSTER, Wuhan, China), p21 (sc-6246, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP9 (10375-2-AP, 1:300, Proteintech, Chicago, IL, USA), MMP2 (10373-2-AP, 1:400, Proteintech), and F4/80 (#70076, 1:200, CST, Boston, MA, USA).

Techniques: Control, Immunohistochemistry, Western Blot, Staining

Journal: European journal of pharmacology

Article Title: Terazosin attenuates abdominal aortic aneurysm formation by downregulating Peg3 expression to inhibit vascular smooth muscle cell apoptosis and senescence.

doi: 10.1016/j.ejphar.2024.176397

Figure Lengend Snippet: Fig. 6. Silencing Peg3 alleviates Ang II-induced cell senescence and apoptosis in MOVAS. MOVAS cells were exposed to 1 μM Ang II with or without Peg3 knockdown for 48 h. (A) Representative western blots and (B) quantification of PEG3, MMP2, p53, cleaved caspase-3 p21, p16 and p-H2AX protein expressions (n = 3). (C) Relative mRNA levels of Peg3 analyzed by RT-qPCR (n = 6). (D) Relative mRNA levels of Mmp2, Mmp9, p53, p21 and p16 analyzed by RT-qPCR (n = 6). (E) (Left) Representative flow-cytometric plots of Annexin V-APC/7-AAD staining. (Right) Quantification of early apoptosis (Annexin V(+)/7-AAD(−)) and total apoptosis (Annexin V(+)) percentages (n = 6). (F) (Left) Representative images of SA-β-gal staining in MOVAS (scale bar, 50 μm) and (Right) quantification of SA- β-gal positive cells (n = 3). (E) Relative mRNA levels of SASP factors including Il-6, Il-1β, Ccl2, Ccl7, Cxcl1, Cxcl10 and Cxcl12 analyzed by RT-qPCR (n = 3). *p < 0.05 (si-Peg3 group vs NC group) or (si-Peg3+Ang II group vs NC + Ang II group), #p < 0.05 (NC + Ang II group vs NC group) or (si-Peg3+Ang II group vs si-Peg3 group).

Article Snippet: Primary antibodies used included: PEG3 (AF9152, 1:150, Affinity, China), α-SMA (BM0002, 1:200, BOSTER, Wuhan, China), p21 (sc-6246, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP9 (10375-2-AP, 1:300, Proteintech, Chicago, IL, USA), MMP2 (10373-2-AP, 1:400, Proteintech), and F4/80 (#70076, 1:200, CST, Boston, MA, USA).

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, Staining

Journal: European journal of pharmacology

Article Title: Terazosin attenuates abdominal aortic aneurysm formation by downregulating Peg3 expression to inhibit vascular smooth muscle cell apoptosis and senescence.

doi: 10.1016/j.ejphar.2024.176397

Figure Lengend Snippet: Fig. 7. Schematic illustration shows the mechanisms of TZ inhibition on AAA formation. Stress induces the expression of Peg3 in VSMCs, triggering VSMCs senescence, apoptosis, and ECM degradation, eventually leading to AAA formation Treatment with low-dose TZ reduces Peg3 expression, reversing these effects. The picture was drawn by Figdraw. VSMCs, vascular smooth muscle cells. SASP, senescence-associated secretory phenotype. ECM, extracellular matrix.

Article Snippet: Primary antibodies used included: PEG3 (AF9152, 1:150, Affinity, China), α-SMA (BM0002, 1:200, BOSTER, Wuhan, China), p21 (sc-6246, 1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP9 (10375-2-AP, 1:300, Proteintech, Chicago, IL, USA), MMP2 (10373-2-AP, 1:400, Proteintech), and F4/80 (#70076, 1:200, CST, Boston, MA, USA).

Techniques: Inhibition, Expressing

Journal: Journal of Biological Chemistry

Article Title: Endorepellin Evokes Autophagy in Endothelial Cells

doi: 10.1074/jbc.m114.556530

Figure Lengend Snippet: FIGURE 6. Endorepellin induces autophagy through a VEGFR2-depen- dent pathway and evokes transcription of BECN1 gene. A, representative immunoblotting following a 6-h treatment with endorepellin (200 nM) of HUVEC treated with scrambled siRNA (siScr), siRNA targeting the VEGFR2 (siVEGFR2), or SU5416 as indicated. The blotting is representative of three independent experiments with similar results. B, representative luciferase reporter assays of PAE-VEGFR2 BECN1-Luc cells treated with soluble endorepel- lin (200 nM) for the designated times, in the presence or absence of SU5416 (30 M). Data are mean S.E., normalized to total cell protein. The values at 4 and 6 h are statistically significant (***, p 0.001) as compared with time 0 and the SU5416-treated samples.

Article Snippet: JUNE 6, 2014 • VOLUME 289 • NUMBER 23 JOURNAL OF BIOLOGICAL CHEMISTRY 16115 at FU B E R L IN /B IB L IO T H E K C H E M IE on M ay 6, 2015 http://w w w .jbc.org/ D ow nloaded from three separate and validated siRNA oligonucleotides specific for PEG3 (sc-97350) from Santa Cruz Biotechnology (Santa Cruz, CA) or targeting VEGFR2 (s7823) from Invitrogen.

Techniques: Western Blot, Luciferase

Journal: Journal of the American Heart Association

Article Title: Platelet‐Derived Growth Factor Receptor Type α Activation Drives Pulmonary Vascular Remodeling Via Progenitor Cell Proliferation and Induces Pulmonary Hypertension

doi: 10.1161/jaha.121.023021

Figure Lengend Snippet: Figure 8. Lung fibrosis is not induced by PDGFRα activation and is not reduced by PDGFRα inhibition. A through C, Fibrosis evaluation in noninduced (n=6–7) or tamoxifen-induced PW1nLacz/+/PDGFRα+/(S)K/Rosa-CRE+ mice 5 weeks after tamoxifen induction (n=5–6). D to F, Fibrosis evaluation in untreated mice under normoxia (n=5) or IgG and APA5-treated mice after 21 days of CH (n=7–8). A and D, RT-qPCR measurements of mRNA expression for collagen 1a1, collagen 3a1, and transforming growth factor-β in lungs. B and E, Representative images of picrosirius red staining for collagen (red) in pulmonary parenchyma. Larger images are displayed in Figures S10 and S11. C and F, Quantification of the picrosirius red-stained area as a measure of fibrosis using Histolab analysis. Fibrosis area (% of total area) was determined as the mean fibrotic area of four large lung sections for each animal. Bars represent means and whiskers represent SD. *P<0.05, **P<0.01 Tam+ vs Tam- group, 2-tailed Mann-Whitney; #P<0.05, ##P<0.01 vs normoxic, Kruskal-Wallis and Dunn. CH indicates chronic hypoxia; ns, not significant for APA5 vs IgG within CH 21d group in D and F (scale bar 200 µm); PDGFRα, platelet-derived growth factor receptor type α; PW1, protein widely 1; and RT-qPCR, reverse transcriptase quantitative polymerase chain reaction.

Article Snippet: Target antigen Provider, reference Dilution, conditions Secondary antibody Provider, reference Dilution, conditions PW1 (human) Novus Biologicals NBP2-46379 1/100 ON; 4°C Alexa Fluor Donkey anti-mouse 647 Thermo Fisher Scientific A31571 1/200 1h ; RT PDGFRα (human) abcam ab61219 1/200 ON; 4°C Alexa Fluor Donkey anti-rabbit 547 Thermo Fisher Scientific A10040 1/200 1h ; RT D ow nloaded from http://ahajournals.org by on O ctober 7, 2024 PW1 (mouse) Relaix 1996 ref 49 1/5000 ON; 4°C Goat anti-rabbit 488 Thermo Fisher Scientific A11034 1/500 1h; RT β-galactosidase Abcam ab9361 1/6000 ON; 4°C Goat anti-chicken 594 Thermo Fisher Scientific A11042 1/500 1h; RT α-SMA-FITC Sigma F3777 1/500 1h30; RT α-SMA ebioscience 14-9760-82 1/500 ON; 4°C Goat anti-mouse 555 Abcam Ab150078 1/500 1h; RT Von Willebrand Factor DAKO A0082 1/300 ON; 4°C Goat anti-rabbit 488 Thermo Fisher Scientific A11034 1/500 1h; RT Ki67 Abcam ab15580 1/300 ON; 4°C Goat anti rabbit-488 Thermo Fisher Scientific A11034 1/500 1h; RT DAPI Santa Cruz sc3598 1/1000 1h; RT

Techniques: Activation Assay, Inhibition, Quantitative RT-PCR, Expressing, Staining, MANN-WHITNEY, Derivative Assay, Reverse Transcription, Real-time Polymerase Chain Reaction