Anti tfe3

For immunofluorescence staining of BMDM or sorted cells, fixed cells were permeabilized with 0.1% Triton X-100/PBS for 10 min and blocked with 5% goat serum/PBS for 30min. Cells were then incubated with 1:500 primary antibody in 5% goat serum/PBS for 2 h at RT. Primary antibodies used were: anti-TFEB (Bethyl Laboratories, A303-673A), anti-TFE3 (Sigma, HPA023881), anti-LAMP1 (Invitrogen, 14-1071-82), anti-ATP6V0D2 (Sigma, SAB2103221, verified using isotype control), anti- ATP6V1B2 antibody (Santa Cruz, sc-166122, verified using isotype control) and anti-CTSB (Abcam, ab58802). Then cells were washed three times with PBS and stained with 1:1000 secondary antibody tagged with Alexa Fluor 647 (see S1 Table). Samples were washed three times with PBS and mounted ProLong Diamond Antifade Mountant with DAPI.

Paraffin-embedded sections were firstly deparaffinized and then incubated with rabbit polyclonal anti-TFE3 (Sigma-Aldrich) primary antibody at 4 °C overnight. After three times wash by PBST, the sections were then incubation with HRP conjugated goat anti-mouse or goat anti-rabbit secondary antibody. The sections were washed by PBST for three times, and the signal was detected using DAB Substrate Kit following the manufacture’s instruction.