Immunofluorescence Staining of Cryosections

Sections were stained with the primary antibodies: anti-human CD105 (endoglin, ENG) biotin-conjugated (1:100, BioLegend, 323214), anti-human CD31-Alexa Fluor 594 conjugated (1:400, BioLegend, 303126), and rabbit anti‐laminin for two hours at room temperature. After PBST washing, the slides were incubated with streptavidin-Alexa Fluor 647 conjugated (1:500, Life Technologies, S21374) and goat anti‐rabbit Alexa Fluor 750-conjugated for an hour. After final PSBT washing, nuclei were counterstained with 4’,6-diamidino-2-phenylindole (DAPI) (0.5 μg/mL, Sigma-Aldrich) and were mounted with ProLong Gold antifade reagent. Cryosections were imaged with Axio Scan.Z1 slide scanner.

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Abbassi-Daloii T., el Abdellaoui S., Voortman L.M., Veeger T.T., Cats D., Mei H., Meuffels D.E., van Arkel E., 't Hoen P.A., Kan H.E, & Raz V. (2023). A transcriptome atlas of leg muscles from healthy human volunteers reveals molecular and cellular signatures associated with muscle location. eLife, 12, e80500.

Publication 2023

anti-human CD105 4’,6-diamidino-2-phenylindole (DAPI)

Alexa 594 Alexa fluor 647 Alexa fluor 750 Antibodies Biotin Cryosections Endoglin Goat Gold Human Laminin Nuclei Rabbit Scan Streptavidin

Corresponding Organization : Maastricht University

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Variable analysis

independent variables

Primary antibodies:
- anti-human CD105 (endoglin, ENG) biotin-conjugated (1:100, BioLegend, 323214)
- anti-human CD31-Alexa Fluor 594 conjugated (1:400, BioLegend, 303126)
- rabbit anti‐laminin

dependent variables

Not explicitly mentioned

control variables

Incubation time: two hours at room temperature
PBST washing
Incubation with streptavidin-Alexa Fluor 647 conjugated (1:500, Life Technologies, S21374) and goat anti‐rabbit Alexa Fluor 750-conjugated for an hour
Final PBST washing
Nuclei counterstained with 4',6-diamidino-2-phenylindole (DAPI) (0.5 μg/mL, Sigma-Aldrich)
Mounted with ProLong Gold antifade reagent
Cryosections imaged with Axio Scan.Z1 slide scanner

controls

Positive control: Not specified
Negative control: Not specified

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