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. 2020 Apr 1;9:e51576. doi: 10.7554/eLife.51576

Figure 2. Validation of satellite cell clusters.

(a) Feature plots displaying localized gene expression for the genes DLK1, ICAM1, and VCAM1 within the 2D UMAP space of the eight combined vasti, the rectus femoris, the two combined rectus abdominis and the two combined pectoralis majors as shown in Figure 1b and Figure 1—figure supplement 2b, Figure 2—figure supplement 1a,b. Each dot represents a single cell. Deeper purple coloration represents increased expression. (b) Immunofluorescence staining of human satellite cells for validation markers within sections of human muscle. Pink arrows denote human satellite cells that are positive for the marker (scale, 50 µm). White arrowheads mark satellite cells that are negative for expression of the marker. (n = 3, biological replicates). (c) Bar plot displaying the quantification of DLK1, ICAM1, and VCAM1 expression of human satellite cells with both immunofluorescence staining and flow cytometry. (n = 3, biological replicates). Data presented as mean ± SEM.

Figure 2—source data 1. Validation of satellite cell clusters.

Figure 2.

Figure 2—figure supplement 1. Validation of satellite cell cluster marker CYCS across multiple human muscle type.

Figure 2—figure supplement 1.

(a-b) UMAP plot of cells isolated from two recti abdominis and two pectoralis major. (c) Feature plot depicting localized gene expression of cluster marker CYCS in the UMAP 2D space in the eight vasti lateralis, rectus femoris, two recti abdominis and two pectoralis major. (d) Immunofluorescence staining of human satellite cells for CYCS within sections of human muscle (scale, 50 µm). Lavender arrows denote human satellite cells that are positive for CYCS. White arrowheads mark satellite cells that are negative for CYCS (n = 3, biological replicates). (e) Bar plot displaying the quantification CYCS expression of PAX7+ human satellite cells by immunofluorescence staining (n = 3, biological replicates). Data presented as mean ± SEM.
Figure 2—figure supplement 1—source data 1. Validation of satellite cell cluster marker CYCS across multiple human muscle type.
Figure 2—figure supplement 2. Heterogeneous expression of ICAM1, VCAM1 and DLK1 in human satellite cells.

Figure 2—figure supplement 2.

(a-c) Representative flow cytometry profiles for CXCR4+/CD29+/CD56+ human satellite cells expressing (a) ICAM1, (b) VCAM1, and (c) DLK1. (n = 3, biological replicates). (d) Representative immunofluorescence co-staining of PAX7+ human satellite cells for DLK1/ICAM, VCAM1/ICAM1 and VCAM1/DLK1 (scale, 50 µm). Lavender arrows mark double positive PAX7+ cells, white arrowheads denote single positive PAX7+ cells. (n = 3, biological replicates). (e) Bar graph showing the proportion of double and single positive satellite cells for DLK1, VCAM1 and ICAM1 in human muscle as quantified by immunofluorescence staining (n = 3, biological replicates). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Data presented as mean ± SEM.