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: International journal of biological sciences

Article Title: FGF7 and FGF10 Promote Fate Transition of Human Epidermal Cell-derived Organoids to an Eccrine Gland Phenotype.

doi: 10.7150/ijbs.97422

Figure Lengend Snippet: Figure 6. Addition of inhibitors targeting MEK1/2 impairs reconstitution of FDKCs into ESG-characterized organoids. (A) Phase-contrast images show the morphology of 3D organoids constructed by "FDKCs + Matrigel" in the presence of FGF7 and FGF10 with or without the MEK1/2 inhibitor U0126 addition at day 9 of culture. (B) RT-qPCR detection of transcriptional expression levels of ERK1/2, FGFR1, FGFR2, K7, and K73 in FGF-fed organoids with (+) or without (-) the addition of the MEK1/2 inhibitor U0126. (C, D) Protein expression of ERK1/2, p-ERK1/2, FGFR1, FGFR2, K7, and K73 in FGF- fed organoids with (+) or without (-) the addition of the MEK1/2 inhibitor U0126. GAPDH serves as the internal control. (C) WB assay. (D) The relative integrated density of ERK1/2, p-ERK1/2, FGFR1, FGFR2, K7, and K73 in accordance with (C). The asterisk (*) indicate significant differences at p < 0.05, and the asterisks (**) indicate significant differences at p < 0.001 (one-way ANOVA, t test).

Article Snippet: For the pathway interference assay experiment, 40 μM MEK1/2 specific inhibitor U0126 (S1901, Beyotime, Shanghai, China), 50 μM FGFR1 inhibitor PD166866 (S8493; Selleck,) and 50 μM FGFR2 inhibitor Formononetin (HY-N0183; MCE) were added to KSFM complete medium in the presence or absence of human FGF7 and FGF10, respectively.

Techniques: Construct, Quantitative RT-PCR, Expressing, Control