Mouse anti mmp1 3a6b4

Staining was carried out using standard protocols. Briefly, wing discs from third instar larva were dissected and fixed in PBS‐T containing 4% formaldehyde for 20 minutes at room temperature. After washing with PBS‐T, they were blocked with 5% BSA in PBS‐T for 30 minutes and then incubated with primary antibodies overnight at 4°C. Washed discs were subsequently incubated with secondary antibodies. The following primary antibodies were used: mouse anti‐MMP1 (3A6B4, 1:300) and mouse anti‐Wg (4D4, 1:500) were purchased from Developmental Studies Hybridoma Bank. Mouse anti‐β‐galactosidase (sc‐65670, 1:500) was from Santa Cruz. Rabbit anti‐Caspase3 (#9661, 1:1000) and rabbit anti‐p‐Histone H3 (#9701, 1:1000) was from Cell Signaling Technology. Secondary antibodies goat anti‐mouse Alexa Fluor594 (A11012, 1:1000) and goat anti‐rabbit Alexa Fluor594 (A11005, 1:1000) were from Invitrogen. Fluorescence images were recorded using a Nikon DS‐Ri1 fluorescence microscope and a Zeiss LSM 880 confocal microscope. Images of the adult eyes were acquired using a Nikon SMZ‐745T trinocular stereo microscope. Images were then analysed using Zeiss Zen, Image J and Adobe Photoshop software.

Antibody staining was performed according to standard procedures. The following primary antibodies were used: mouse anti-MMP1 (3A6B4, 1:200, Developmental Studies Hybridoma Bank, DSHB), Rat anti-E-cad (DCAD2-c, 1:100, Developmental Studies Hybridoma Bank, DSHB) and mouse anti $\beta$ -gal (40-1a, 1:500, Developmental Studies Hybridoma Bank, DSHB). The following secondary antibodies were used: anti-mouse CY3 (A11032, 1:1000, Cell Signaling Technology) and anti-Rat CY3 (104,086, 1:500, Jackson Immuno research).