Anti tgf beta 1

Primary antibodies used for immunoblotting and immunostaining: anti-annexin A6 (cat# ab199422; Abcam), anti-annexin A2 (cat# 610068; BD Bioscience), anti-caveolin 3 (cat# NBP128917; Novus Bio), anti-collagen type 1 (cat# CL50151AP1; Cedarlane), anti-decorin (cat# AF1060; R&D Systems), anti-dystrophin (cat# PA137587; Invitrogen), anti-HSP90 (cat#4874; Cell Signaling Technologies), anti-laminin-2 (α–2 Chain) (cat# L0663; MilliporeSigma), anti-MMP9 (cat# ab38898; Abcam), anti-myosin 4 (cat# PA550065; Invitrogen), anti-periostin/OSF-2 (cat# NBP130042; Novus Bio), anti-Mo CD140a (PDGRFα) (cat# 17140181; Invitrogen), anti-TGF Beta 1 (cat# 218981AP; Proteintech and cat# MA515065; Invitrogen), anti-Thrombospondin-4 (cat# AF2390; R&D Systems), and anti-TIMP-1 (cat# AF980; R&D Systems). Secondary antibodies used: Alexa Fluor 488 goat anti-rat (cat# A11006; Invitrogen), Alexa Fluor 594 goat anti-rabbit (cat# A11012; Invitrogen) and Alexa Fluor 594 donkey anti-goat (cat# A11058; Invitrogen).

Decellularized muscle cryosections (dECMs) were fixed in 4% paraformaldehyde, rinsed, and blocked with a 1% BSA/10% FBS blocking buffer at room temperature for 1 hour. Scaffolds were incubated in primary antibodies overnight at 4°C at 1:100: anti-laminin-2 (α-2 Chain) (cat# L0663; MilliporeSigma), anti-TGF Beta 1 (cat# 218981AP; Proteintech and cat# MA515065; Invitrogen), anti-Thrombospondin-4 (cat# AF2390; R&D Systems), anti- TGF-β3 (cat# ab15537; Abcam). Secondary antibodies were used at 1:2,500 for 1 hr at room temperature: Alexa Fluor 488 goat anti-rat (cat# A11006; Invitrogen), Alexa Fluor 594 goat anti-rabbit (cat# A11012; Invitrogen) and Alexa Fluor 594 donkey anti-goat (cat# A11058; Invitrogen). The dECMs were mounted with ProLong Gold Antifade Reagent (cat# P36934; Invitrogen). Images were acquired on a Keyence BZ-X810 microscope with identical exposure settings were across genotypes.