mouse pulmonary artery smooth muscle cells (pasmcs) Search Results


mouse pulmonary artery smooth muscle cells (pasmcs)  (Procell Inc)
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Procell Inc mouse pulmonary artery smooth muscle cells (pasmcs)
CircMyst4 is downregulated in PH (A) Genomic location of circRNA Myst4 (circMyst4). CircMyst4 was formed by the back-splicing of exons 2 of Myst4. And the back-splicing junction site of circMyst4 was identified by Sanger sequencing. (B) Real-time qPCR analysis of genomic DNA (gDNA) and cDNA using divergent and convergent primers of circMyst4 ( n = 6). (C) RNase R degradation analysis was performed to detect the stability of circMyst4 and linear Myst4 mRNA ( n = 6). (D) Real-time qPCR analysis was conducted to detect the amount of circMyst4 and linear Myst4 mRNA in <t>PASMCs</t> after actinomycin (D) treatment ( n = 6). (E and F) Real-time qPCR analysis of circMyst4 expression levels in lung tissues of hypoxic PH mouse ( n = 8 mice/group) and hypoxic PASMCs ( n = 6). (G) Real-time qPCR analysis of circMyst4 expression levels in plasma of hypoxic PH mouse and normoxic mouse ( n = 10). (H) Fluorescence in situ hybridization analysis of circMyst4 location in lung tissues of mouse. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI <t>(blue),</t> <t>pulmonary</t> smooth muscle stained with α-SMA (green). Scale bar, 100 μm ( n = 6 mice/group). (I) Real-time qPCR analysis was using to determine the circMyst4 expression in the nucleus and cytoplasm of PASMCs after exposure to hypoxia for 24 h ( n = 6). (J) RNA FISH analysis of the subcellular localization of circMyst4 in PASMCs. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI (blue). U6 and 18S RNA, used as internal references, were labeled with Cy3 (red). Scale bar, 100 μm ( n = 6). Data are shown as means ± SD. Statistical analysis was performed with Student’s t test. Hyp, hypoxia; Nor, normoxia; ns, not significantly different. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.
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CircMyst4 is downregulated in PH (A) Genomic location of circRNA Myst4 (circMyst4). CircMyst4 was formed by the back-splicing of exons 2 of Myst4. And the back-splicing junction site of circMyst4 was identified by Sanger sequencing. (B) Real-time qPCR analysis of genomic DNA (gDNA) and cDNA using divergent and convergent primers of circMyst4 ( n = 6). (C) RNase R degradation analysis was performed to detect the stability of circMyst4 and linear Myst4 mRNA ( n = 6). (D) Real-time qPCR analysis was conducted to detect the amount of circMyst4 and linear Myst4 mRNA in PASMCs after actinomycin (D) treatment ( n = 6). (E and F) Real-time qPCR analysis of circMyst4 expression levels in lung tissues of hypoxic PH mouse ( n = 8 mice/group) and hypoxic PASMCs ( n = 6). (G) Real-time qPCR analysis of circMyst4 expression levels in plasma of hypoxic PH mouse and normoxic mouse ( n = 10). (H) Fluorescence in situ hybridization analysis of circMyst4 location in lung tissues of mouse. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI (blue), pulmonary smooth muscle stained with α-SMA (green). Scale bar, 100 μm ( n = 6 mice/group). (I) Real-time qPCR analysis was using to determine the circMyst4 expression in the nucleus and cytoplasm of PASMCs after exposure to hypoxia for 24 h ( n = 6). (J) RNA FISH analysis of the subcellular localization of circMyst4 in PASMCs. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI (blue). U6 and 18S RNA, used as internal references, were labeled with Cy3 (red). Scale bar, 100 μm ( n = 6). Data are shown as means ± SD. Statistical analysis was performed with Student’s t test. Hyp, hypoxia; Nor, normoxia; ns, not significantly different. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Journal: iScience

Article Title: Superenhancer-driven circRNA Myst4 involves in pulmonary artery smooth muscle cell ferroptosis in pulmonary hypertension

doi: 10.1016/j.isci.2024.110900

Figure Lengend Snippet: CircMyst4 is downregulated in PH (A) Genomic location of circRNA Myst4 (circMyst4). CircMyst4 was formed by the back-splicing of exons 2 of Myst4. And the back-splicing junction site of circMyst4 was identified by Sanger sequencing. (B) Real-time qPCR analysis of genomic DNA (gDNA) and cDNA using divergent and convergent primers of circMyst4 ( n = 6). (C) RNase R degradation analysis was performed to detect the stability of circMyst4 and linear Myst4 mRNA ( n = 6). (D) Real-time qPCR analysis was conducted to detect the amount of circMyst4 and linear Myst4 mRNA in PASMCs after actinomycin (D) treatment ( n = 6). (E and F) Real-time qPCR analysis of circMyst4 expression levels in lung tissues of hypoxic PH mouse ( n = 8 mice/group) and hypoxic PASMCs ( n = 6). (G) Real-time qPCR analysis of circMyst4 expression levels in plasma of hypoxic PH mouse and normoxic mouse ( n = 10). (H) Fluorescence in situ hybridization analysis of circMyst4 location in lung tissues of mouse. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI (blue), pulmonary smooth muscle stained with α-SMA (green). Scale bar, 100 μm ( n = 6 mice/group). (I) Real-time qPCR analysis was using to determine the circMyst4 expression in the nucleus and cytoplasm of PASMCs after exposure to hypoxia for 24 h ( n = 6). (J) RNA FISH analysis of the subcellular localization of circMyst4 in PASMCs. CircMyst4 probes were labeled with Cy3 (red). Nucleus were stained with DAPI (blue). U6 and 18S RNA, used as internal references, were labeled with Cy3 (red). Scale bar, 100 μm ( n = 6). Data are shown as means ± SD. Statistical analysis was performed with Student’s t test. Hyp, hypoxia; Nor, normoxia; ns, not significantly different. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Mouse pulmonary artery smooth muscle cells (PASMCs) were obtained from Procell Life Science & Technology (Wuhan, China), and were cultured in Dulbecco’s Modified Eagle Medium (DMEM) containing 15% fetal bovine serum (FBS) and 1% penicillin/streptomycin (PEST) and were placed in a 37°C humidified incubator containing 5% CO2.

Techniques: Sequencing, Expressing, Clinical Proteomics, Fluorescence, In Situ Hybridization, Labeling, Staining