Figure 1. Expression of SR Ca 2 channels... | Rockefeller University Press

Figure 1.

Expression of SR Ca2+channels in IALVs lymphatic muscle cells. (A) Lymphatic muscle cells in the Myh11Cre-EGFP mouse express EGFP and wrap circumferentially around the vessel wall. The scale bar is 100 µm. (B) Digestion of the IALVs from Myh11Cre-EGFP mice into single cells and sorting of the GFP+ lymphatic muscle population. (C and D) Gel electrophoresis of the RT-PCR of the SR Ca2+ channels in (C) whole IALVs and (D) sorted LMCs from B. (E) FACs-purified LMC identity was confirmed via RT-PCR for the lymphatic muscle cell markers Myh11 and calponin (Cnn1) and lack of expression of the lymphatic endothelial cell marker Prox1 and the fibroblast marker PDGFRα. FACs and RT-PCR were conducted in three independent experiments with four mice. For two of the experiments, both the left and right IALV were pooled from a single male mouse while a third experiment included both IALVs pooled from two female mice. (F) Small intestine and skeletal muscle served as positive controls for the primers. Each image is a single gel. # indicates apparent primer dimer presence. Source data are available for this figure: SourceData F1.

Expression of SR Ca ²⁺ channels in IALVs lymphatic muscle cells. (A) Lymphatic muscle cells in the Myh11Cre-EGFP mouse express EGFP and wrap circumferentially around the vessel wall. The scale bar is 100 µm. (B) Digestion of the IALVs from Myh11Cre-EGFP mice into single cells and sorting of the GFP⁺ lymphatic muscle population. (C and D) Gel electrophoresis of the RT-PCR of the SR Ca²⁺ channels in (C) whole IALVs and (D) sorted LMCs from B. (E) FACs-purified LMC identity was confirmed via RT-PCR for the lymphatic muscle cell markers Myh11 and calponin (Cnn1) and lack of expression of the lymphatic endothelial cell marker Prox1 and the fibroblast marker PDGFRα. FACs and RT-PCR were conducted in three independent experiments with four mice. For two of the experiments, both the left and right IALV were pooled from a single male mouse while a third experiment included both IALVs pooled from two female mice. (F) Small intestine and skeletal muscle served as positive controls for the primers. Each image is a single gel. # indicates apparent primer dimer presence. Source data are available for this figure: SourceData F1.