Journal: International Journal of Molecular Sciences

Article Title: PIEZO1 Mediates Apoptosis of Endothelial Cells via Enhancing HMGA2 Expression Under Simulated Microgravity

doi: 10.3390/ijms27031425

Figure Lengend Snippet: Simulated microgravity promotes PIEZO1 expression in HUVECs. ( A ) qRT-PCR analysis of PIEZO1 in HUVECs after simulated microgravity for 48 h. ( B ) Western blot analysis for the expression of PIEZO1 in HUVECs after simulated microgravity for 48 h. ( C ) The immunofluorescence staining of PIEZO1 (red immunofluorescence) and DAPI (blue immunofluorescence). (The data represent the mean ± SEM , n = 3; ns (no significance), p > 0.05; * p < 0.05, Scale Bar = 50 μm).

Article Snippet: Subsequently, the cells were incubated overnight at 4 °C with a PIEZO1 antibody (1:200; Proteintech, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: PIEZO1 Mediates Apoptosis of Endothelial Cells via Enhancing HMGA2 Expression Under Simulated Microgravity

doi: 10.3390/ijms27031425

Figure Lengend Snippet: Simulated microgravity triggers endothelial cell apoptosis through PIEZO1 activation. ( A ) The knockdown efficiency of PIEZO1 in HUVECs was validated by qRT-PCR. ( B ) PIEZO1 protein knockdown in HUVECs was further confirmed by Western blot. ( C ) Analysis of cell apoptosis in HUVECs using Annexin V and PI staining through flow cytometry. Con+si-NC (control group transfected with negative control siRNA), SMG+si-NC (Simulated microgravity-treated group transfected with negative control siRNA), SMG+si-PIEZO1 (Simulated microgravity-treated group transfected with PIEZO1 siRNA). ( D ) Analysis of apoptosis-related proteins in HUVECs using Western blot. (The data represent the mean ± SEM , n = 3; ns (no significance), p > 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Subsequently, the cells were incubated overnight at 4 °C with a PIEZO1 antibody (1:200; Proteintech, Wuhan, China).

Techniques: Activation Assay, Knockdown, Quantitative RT-PCR, Western Blot, Staining, Flow Cytometry, Control, Transfection, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: PIEZO1 Mediates Apoptosis of Endothelial Cells via Enhancing HMGA2 Expression Under Simulated Microgravity

doi: 10.3390/ijms27031425

Figure Lengend Snippet: The proteomics of HUVECs after knockdown of PIEZO1 under simulated microgravity. ( A , B ) Volcanic maps of differential proteins, where red dots indicate significant upregulation, blue dots indicate significant downregulation, and redindicates no significant differences. ( C ) A Venn diagram compared the significant DEPs of SMG+si-NC vs.Con+si-NC and SMG+si-PIEZO1 vs. SMG+si-NC. ( D ) The GO analysis of DEPs. ( n = 3).

Article Snippet: Subsequently, the cells were incubated overnight at 4 °C with a PIEZO1 antibody (1:200; Proteintech, Wuhan, China).

Techniques: Knockdown

Journal: International Journal of Molecular Sciences

Article Title: PIEZO1 Mediates Apoptosis of Endothelial Cells via Enhancing HMGA2 Expression Under Simulated Microgravity

doi: 10.3390/ijms27031425

Figure Lengend Snippet: PIEZO1 triggers an increase in HMGA2 expression through the influx of Ca 2+ . ( A , B ) Following 48 h of microgravity exposure, the expression of HMGA2 in HUVECs, with or without si-PIEZO1, was analyzed using qRT-PCR and Western blot. ( C ) Confocal microscopy and Immunofluorescence quantification assay were as used to detect Ca 2+ influx. Ca 2+ was stained with green immunofluorescence, while DAPI was stained with blue immunofluorescence. (Scale Bar = 100 μm). ( D ) The expression of HMGA2 in HUVECs, with or without BAPTA, was analyzed using Western blot after 48 h of simulated microgravity exposure. (The data represent the mean ± SEM , n = 3; ns (no significance), p > 0.05; * p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Subsequently, the cells were incubated overnight at 4 °C with a PIEZO1 antibody (1:200; Proteintech, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Confocal Microscopy, Immunofluorescence, Staining