Mouse primary antibody for collagen type 1

For immunofluorescence, CTRL and H10 gels have been prepared as already mentioned. After 24 h, gels have been incubated in PBS 1X with 3% of bovine serum albumin (BSA, Sigma Aldrich, Oakville, ON, Canada) for 10 min. Then gels have been incubated with mouse primary antibody for collagen type 1 (1: 1,000; Novus Biological, Oakville, ON, Canada) for 2 h at 37°C. Following, gels were incubated with an Alexa Fluor® 488 goat anti-mouse secondary antibody (Life Technologies, Sigma Aldrich, Oakville, ON, Canada) for 2 h at room temperature under agitation. Afterwards, gels have been rinsed three time with PBS 1X with 0.01% Tween 20 and have been kept overnight at 4°C before being placed on fluorescent microscope slides. Images at a magnification of 20X have been collected using an Olympus BX51 Fluorescence Microscope (Olympus Canada Inc., Toronto, ON, Canada).

For immunofluorescence, CTRL and H10 gels have been prepared as already mentioned. After 24 h, gels have been incubated in PBS 1X with 3% of bovine serum albumin (BSA, Sigma Aldrich, Oakville, ON, Canada) for 10 min. Then gels have been incubated with mouse primary antibody for collagen type 1 (1:1,000; Novus Biological, Oakville, ON, Canada) for 2 h at 37°C. Following, gels were incubated with an Alexa Fluor® 488 goat anti-mouse secondary antibody (Life Technologies, Sigma Aldrich, Oakville, ON, Canada) for 2 h at room temperature under agitation. Afterwards, gels have been rinsed three time with PBS 1X with 0.01% Tween 20 and have been kept overnight at 4°C before being placed on fluorescent microscope slides. Images at a magnification of 20X have been collected using an Olympus BX51 Fluorescence Microscope (Olympus Canada Inc., Toronto, ON, Canada).