calm4  (Cusabio)

Journal: Materials Today Bio

Article Title: Porcine pericardial decellularized matrix bilayer patch containing adipose stem cell-derived exosomes for the treatment of diabetic wounds

doi: 10.1016/j.mtbio.2024.101398

Figure Lengend Snippet: The exosome-loaded bilayer decellularized pericardial patches promote the healing of diabetic refractory wounds by enhancing the activation of M2 macrophages in the wound and activating the cAMP signaling pathway. (A) On day 7, dual immunofluorescence staining for F4/80 (M0 marker), iNOS (M1 marker), and CD206 (M2 marker) was performed to assess the numbers of activated M1 and M2 macrophages in the wound and surrounding skin tissue, scale bar = 50 μm. (B) On day 7, RT-qPCR was used to detect the expression levels of M1 ( cd86 , il-6 ) and M2 ( cd163 , arg-1 ) macrophage-related genes in the wound tissue of each group. (C) On day 7, Western Blotting experiments were conducted to measure the expression levels of key proteins in the cAMP signaling pathway (Atp1a2, Calm4, and Cngb1) in the wound tissue of each group, with quantitative analysis of protein density, n = 3 (D). ∗, p < 0.05, ∗∗, p < 0.01, and ∗∗∗, p < 0.001, compared with the PBS group.

Article Snippet: The primary antibodies used were as follows: Atp1a2 (ab166888, Abcam, USA), 1:2000; Calm4 (CSB-PA004454GA01HU, Cusabio), 1:1000; Gngb1 (PA5-101543, Invitrogen), 1:1000; GAPDH (#2118S, CST), 1:1000.

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

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 small-interfering RNA (siRNA) inhibits pyroptosis in pulmonary artery smooth muscle cells (PASMCs). A and B , Circ-Calm4 siRNA reversed the increased protein and mRNA levels of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment), IL-18 (interleukin-18), and IL-1β (interleukin-1β) induced by hypoxia in PASMCs (n=6). C , Circ-Calm4 siRNA reversed the increased LDH (lactate dehydrogenase) activity in PASMCs subjected to hypoxia for 24 h. LDH release was evaluated with an LDH release kit (n=6). D , Knockdown of circ-Calm4 decreased the number of pyroptotic cells in PASMCs exposed to hypoxia. Cells were detected with annexin V-FITC/propidium iodide (PI) double staining using quantitative fluorescence-activated cell sorting (FACS) analysis. E , Knockdown of circ-Calm4 by siRNA reduced the positive PI staining induced by hypoxia in PASMCs. Scale bars=100 µm. Images of fluorescence staining with PI (red) and Hoechst 33342 (blue). F , Circ-calm4 siRNA attenuated the fluorescence staining of Caspase-1 and IL-18 induced by hypoxia in PASMCs. Scale bars=100 µm. Cells were stained for Caspase-1 (green) and IL-18 (red), and DAPI (blue) was used for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. The graph A-ASC and Caspase-1 were analyzed by the Kruskal-Wallis test followed by Dunn post-test. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques: Small Interfering RNA, Binding Assay, Activity Assay, Double Staining, Fluorescence, FACS, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 inhibits pyroptosis in a mouse model of hypoxia-induced pulmonary hypertension (PH). A and B , Knockdown of circ-Calm4 by sh-circ-Calm4 countered the hypoxia-induced upregulation of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment ),IL-18 (interleukin-18), and IL-1β (interleukin-1β) at the protein and mRNA levels (n=6; in Western blot, the image of NLPR3 and Caspase-1 used the same actin). C and D , Knockdown of circ-Calm4 by sh-circ-Calm4 reversed the hypoxia-induced upregulation of Caspase-1 and IL-18 in mouse lung tissues. Scale bars=100 µm. Lung sections stained with Caspase-1 (green) and IL-18 (green), pulmonary smooth muscle stained with α-SMA (red), and DAPI for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. The graph B-ASC was analyzed by the Kruskal-Wallis test followed by Dunn post-test. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques: Binding Assay, Western Blot, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-calm4 is a mediator that regulates miR-124-3p action. A , The target microRNAs (miRNAs) of circ-Calm4, as predicted by the miRanda Count, RNA hybrid Count and RegRNA 2.0 databases. B , Knockdown of circ-Calm4 by small-interfering RNA increased the miR-124-3p mRNA level in pulmonary arterial smooth muscle cells (PASMCs) exposed to hypoxia for 24 h (n=6). C , HEK293 cells were cotransfected with a luciferase reporter construct carrying wild-type (WT) or mutant (Mut) circ-Calm4 and miR-124-3p or miR-124-3p-NC. Luciferase activities were measured via a dual luciferase assay (n=5). D , Colocalization of circ-Calm4 and miR-124-3p in PASMCs. Scale bars=100 µm. MiR-124-3p probes were labeled with FAM (green), whereas circ-Calm4 probes were labeled with Cy3 (red). Nuclei were stained with DAPI (blue). Pearson coefficient is 0.77809, indicating a correlation. E , Downregulation of miR-124-3p in PASMCs exposed to hypoxia for 24 h relative to miR-124-3p expression in nontreated cells (n=8). F , Relative fluorescence of miR-124-3p in PASMCs exposed to hypoxia. Scale bars=100 µm. miR-124-3p probes were labeled with FAM (green). Nuclei were stained with DAPI (blue). Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction and Student t test for 2 means. * P< 0.05, **P< 0.01, *** P< 0.001.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques: Small Interfering RNA, Luciferase, Construct, Mutagenesis, Labeling, Staining, Expressing, Fluorescence

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 sponges miR-124-3p to mediate pulmonary arterial smooth muscle cell (PASMC) pyroptosis. A and B , Knockdown of endogenous miR-124-3p by AMO-124-3p abrogated the antipyroptotic effects of Circ-calm4 silencing by Circ-calm4-small-interfering RNA (siRNA) upon hypoxia exposure in PASMCs, as indicated by the protein and mRNA levels of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment), IL-18 (interleukin-18), and IL-1β (interleukin-1β; n=6). C , Cotransfection with circ-Calm4 siRNA and AMO-124-3p abrogated the antipyroptotic effects on PASMCs upon hypoxia exposure, as indicated by LDH (lactate dehydrogenase) activity (n=6). D , Knockdown of circ-Calm4 by siRNA reduced the positive propidium iodide (PI) staining induced by hypoxia in cells, whereas AMO-124-3p reversed the decrease in PI-positive PASMCs. Scale bars=100 µm. Images of fluorescence staining with PI (red) and Hoechst 33342 (blue). E , Circ-Calm4 knockdown blocked the fluorescence intensity of Caspase-1 and IL-18 under hypoxia exposure, and this decrease was abrogated after transfection of AMO-124-3p. Scale bars=100 µm. Cells were stained for Caspase-1 (green) and IL-18 (red), and DAPI (blue) was used for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques: Small Interfering RNA, Binding Assay, Cotransfection, Activity Assay, Staining, Fluorescence, Transfection

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 small-interfering RNA inhibits pyroptosis in pulmonary arterial smooth muscle cells (PASMCs) through the miR-124-3p/PDCD6 (programmed cell death protein 6) pathway. A and B , Knockdown of endogenous circ-Calm4 abrogated upregulation of PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia (n=6). C and D , MiR-124-3p reversed but AMO-124-3p enhanced the upregulation of PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia (n=6). E and F , In PASMCs, knockdown of circ-Calm4 reduced PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia, whereas AMO-124-3p reversed the downregulation of PDCD6 protein and Pdcd6 mRNA expression (n=6). G and H , Knockdown of circ-Calm4 by sh-circ-Calm4 countered the hypoxia-induced upregulation of PDCD6 at the protein and mRNA levels (n=6). I , Circ-Calm4 expression was positively correlated with Pdcd6 mRNA expression in hypoxic mice. In hypoxic mice, circ-Calm4 expression was negatively correlated with miR-124-3p expression, and Pdcd6 mRNA expression was negatively correlated with miR-124-3p expression. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques: Small Interfering RNA, Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: A schematic diagram to illustrate the hypothetical model Circ-Calm4, as a competitive endogenous RNA, adsorbed miR-124-3p via a circular RNA sponging mechanism and abolished the suppression of Pdcd6 (programmed cell death protein 6) by miR-124-3p, leading to pyroptosis, proliferation, and apoptosis resistance. PASMC indicates pulmonary arterial smooth muscle cell.

Article Snippet: Cy3-labeled circ-Calm4 probes and FAM-labeled miR-124-3p probes were designed by GeneChem (Shanghai, China).

Techniques:

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 small-interfering RNA (siRNA) inhibits pyroptosis in pulmonary artery smooth muscle cells (PASMCs). A and B , Circ-Calm4 siRNA reversed the increased protein and mRNA levels of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment), IL-18 (interleukin-18), and IL-1β (interleukin-1β) induced by hypoxia in PASMCs (n=6). C , Circ-Calm4 siRNA reversed the increased LDH (lactate dehydrogenase) activity in PASMCs subjected to hypoxia for 24 h. LDH release was evaluated with an LDH release kit (n=6). D , Knockdown of circ-Calm4 decreased the number of pyroptotic cells in PASMCs exposed to hypoxia. Cells were detected with annexin V-FITC/propidium iodide (PI) double staining using quantitative fluorescence-activated cell sorting (FACS) analysis. E , Knockdown of circ-Calm4 by siRNA reduced the positive PI staining induced by hypoxia in PASMCs. Scale bars=100 µm. Images of fluorescence staining with PI (red) and Hoechst 33342 (blue). F , Circ-calm4 siRNA attenuated the fluorescence staining of Caspase-1 and IL-18 induced by hypoxia in PASMCs. Scale bars=100 µm. Cells were stained for Caspase-1 (green) and IL-18 (red), and DAPI (blue) was used for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. The graph A-ASC and Caspase-1 were analyzed by the Kruskal-Wallis test followed by Dunn post-test. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: Small Interfering RNA, Binding Assay, Activity Assay, Double Staining, Fluorescence, FACS, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 inhibits pyroptosis in a mouse model of hypoxia-induced pulmonary hypertension (PH). A and B , Knockdown of circ-Calm4 by sh-circ-Calm4 countered the hypoxia-induced upregulation of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment ),IL-18 (interleukin-18), and IL-1β (interleukin-1β) at the protein and mRNA levels (n=6; in Western blot, the image of NLPR3 and Caspase-1 used the same actin). C and D , Knockdown of circ-Calm4 by sh-circ-Calm4 reversed the hypoxia-induced upregulation of Caspase-1 and IL-18 in mouse lung tissues. Scale bars=100 µm. Lung sections stained with Caspase-1 (green) and IL-18 (green), pulmonary smooth muscle stained with α-SMA (red), and DAPI for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. The graph B-ASC was analyzed by the Kruskal-Wallis test followed by Dunn post-test. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: Binding Assay, Western Blot, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-calm4 is a mediator that regulates miR-124-3p action. A , The target microRNAs (miRNAs) of circ-Calm4, as predicted by the miRanda Count, RNA hybrid Count and RegRNA 2.0 databases. B , Knockdown of circ-Calm4 by small-interfering RNA increased the miR-124-3p mRNA level in pulmonary arterial smooth muscle cells (PASMCs) exposed to hypoxia for 24 h (n=6). C , HEK293 cells were cotransfected with a luciferase reporter construct carrying wild-type (WT) or mutant (Mut) circ-Calm4 and miR-124-3p or miR-124-3p-NC. Luciferase activities were measured via a dual luciferase assay (n=5). D , Colocalization of circ-Calm4 and miR-124-3p in PASMCs. Scale bars=100 µm. MiR-124-3p probes were labeled with FAM (green), whereas circ-Calm4 probes were labeled with Cy3 (red). Nuclei were stained with DAPI (blue). Pearson coefficient is 0.77809, indicating a correlation. E , Downregulation of miR-124-3p in PASMCs exposed to hypoxia for 24 h relative to miR-124-3p expression in nontreated cells (n=8). F , Relative fluorescence of miR-124-3p in PASMCs exposed to hypoxia. Scale bars=100 µm. miR-124-3p probes were labeled with FAM (green). Nuclei were stained with DAPI (blue). Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction and Student t test for 2 means. * P< 0.05, **P< 0.01, *** P< 0.001.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: Small Interfering RNA, Luciferase, Construct, Mutagenesis, Labeling, Staining, Expressing, Fluorescence

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 sponges miR-124-3p to mediate pulmonary arterial smooth muscle cell (PASMC) pyroptosis. A and B , Knockdown of endogenous miR-124-3p by AMO-124-3p abrogated the antipyroptotic effects of Circ-calm4 silencing by Circ-calm4-small-interfering RNA (siRNA) upon hypoxia exposure in PASMCs, as indicated by the protein and mRNA levels of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment), IL-18 (interleukin-18), and IL-1β (interleukin-1β; n=6). C , Cotransfection with circ-Calm4 siRNA and AMO-124-3p abrogated the antipyroptotic effects on PASMCs upon hypoxia exposure, as indicated by LDH (lactate dehydrogenase) activity (n=6). D , Knockdown of circ-Calm4 by siRNA reduced the positive propidium iodide (PI) staining induced by hypoxia in cells, whereas AMO-124-3p reversed the decrease in PI-positive PASMCs. Scale bars=100 µm. Images of fluorescence staining with PI (red) and Hoechst 33342 (blue). E , Circ-Calm4 knockdown blocked the fluorescence intensity of Caspase-1 and IL-18 under hypoxia exposure, and this decrease was abrogated after transfection of AMO-124-3p. Scale bars=100 µm. Cells were stained for Caspase-1 (green) and IL-18 (red), and DAPI (blue) was used for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: Small Interfering RNA, Binding Assay, Cotransfection, Activity Assay, Staining, Fluorescence, Transfection

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: Circ-Calm4 small-interfering RNA inhibits pyroptosis in pulmonary arterial smooth muscle cells (PASMCs) through the miR-124-3p/PDCD6 (programmed cell death protein 6) pathway. A and B , Knockdown of endogenous circ-Calm4 abrogated upregulation of PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia (n=6). C and D , MiR-124-3p reversed but AMO-124-3p enhanced the upregulation of PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia (n=6). E and F , In PASMCs, knockdown of circ-Calm4 reduced PDCD6 protein and Pdcd6 mRNA expression induced by hypoxia, whereas AMO-124-3p reversed the downregulation of PDCD6 protein and Pdcd6 mRNA expression (n=6). G and H , Knockdown of circ-Calm4 by sh-circ-Calm4 countered the hypoxia-induced upregulation of PDCD6 at the protein and mRNA levels (n=6). I , Circ-Calm4 expression was positively correlated with Pdcd6 mRNA expression in hypoxic mice. In hypoxic mice, circ-Calm4 expression was negatively correlated with miR-124-3p expression, and Pdcd6 mRNA expression was negatively correlated with miR-124-3p expression. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: Small Interfering RNA, Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Circular RNA Calm4 Regulates Hypoxia-Induced Pulmonary Arterial Smooth Muscle Cells Pyroptosis via the Circ-Calm4/miR-124-3p/PDCD6 Axis

doi: 10.1161/ATVBAHA.120.315525

Figure Lengend Snippet: A schematic diagram to illustrate the hypothetical model Circ-Calm4, as a competitive endogenous RNA, adsorbed miR-124-3p via a circular RNA sponging mechanism and abolished the suppression of Pdcd6 (programmed cell death protein 6) by miR-124-3p, leading to pyroptosis, proliferation, and apoptosis resistance. PASMC indicates pulmonary arterial smooth muscle cell.

Article Snippet: The RNA complex was pulled down by incubating the cell lysates with Biotin-labeled circ-calm4 probes (GeneChem Shanghai, China) at 37 °C for 3 hours and streptavidin beads for 30 minutes.

Techniques: