smooth muscle myosin heavy chain  (Grainger Industrial)

Journal: Experimental lung research

Article Title: Store-operated Ca 2+ entry contributes to the ASM phenotype transition in asthma.

doi: 10.1080/01902148.2025.2486951

Figure Lengend Snippet: Figure 1. PDGF-BB induces phenotype switching in ASMCs. A) ASMCs were treated with PDGF-BB (0–50 ng/ml) for 24h. Cell pro liferation rates were measured by the CCK-8 assay and the density was measured by a microplate reader. B) PCNA immunocyto chemical staining in PDGF-BB-treated ASMCs. Scale bar = 100 μm. C) Quantitative analysis of the percentage of PCNA+ ASMCs in 500 cells. D, E, F) the secretion of IL-1β, IL-6, and CXCL-1 from PDGF-BB-treated ASMCs was measured by ELISA. The experiment was repeated three times. G) the expressions of contractile proteins including α-SMA, Sm22α and sm-MHC as well as ECM proteins including collagen I and fibronectin was detected by Western blot. H, I, J) The density quantification of α-SMA, sm22α and sm-MHC was expressed as a ratio relative to GAPDH. K, L) The density quantification of collagen I and fibronectin were expressed as a ratio relative to GAPDH. Multiple groups were analyzed by one-way ANOVA, and Tukey’s HSD test was used for the post-hoc analysis. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group. #p < 0.05, ##p < 0.01, vs. 20 ng/ml PDGF-BB-treated group.

Article Snippet: The primary antibodies of STIM1 (11565-1-AP, 1:1000), Orai1 (28411-1-AP, 1:1000), α-smooth muscle actin (α-SMA) (14395-1-AP, 1:1000), GAPDH (60004-1-Ig, 1:10,000), collagen I (66761-1- Ig, 1:1000) and smooth muscle-myosin heavy chain (sm-MHC) (21404-1-AP, 1:1000) were purchased from Proteintech.

Techniques: CCK-8 Assay, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Experimental lung research

Article Title: Store-operated Ca 2+ entry contributes to the ASM phenotype transition in asthma.

doi: 10.1080/01902148.2025.2486951

Figure Lengend Snippet: Figure 5. SOCE Inhibitors attenuated PDGF-BB-induced reduced contractile protein contents and elevated ECM proteins production in ASMC. ASMCs were treated with PDGF-BB (0, 20, 50 ng/ml) to induce the phenotype switching and several of some were treated in combination with SKF-96365 (15 μM) or RO2959 (10 μM). A) the expression of contractile protein including α-SMA, sm22α, and sm-MHC as well as ECM proteins including fibronectin, collagen-I and matrix metalloproteases were detected by Western blot. B, C, D) The density quantification of α-SMA, sm22α and sm-MHC was expressed as a ratio relative to GAPDH. E–H) The expressions of ECM protein including fibronectin, collagen-I and matrix metalloprotease including MMP2 and MMP9 were detected by Western blot. The experiment was repeated three times. Multiple groups were analyzed by one-way ANOVA, and Tukey’s HSD test was used for the post-hoc analysis. *p < 0.05, **p < 0.01 vs. PDGF-BB 20 ng/ml group. #p < 0.05, ##p < 0.01 vs PDGF-BB 50 ng/ml group.

Article Snippet: The primary antibodies of STIM1 (11565-1-AP, 1:1000), Orai1 (28411-1-AP, 1:1000), α-smooth muscle actin (α-SMA) (14395-1-AP, 1:1000), GAPDH (60004-1-Ig, 1:10,000), collagen I (66761-1- Ig, 1:1000) and smooth muscle-myosin heavy chain (sm-MHC) (21404-1-AP, 1:1000) were purchased from Proteintech.

Techniques: Expressing, Western Blot