Anti calr

The primary antibodies used were β‐actin [anti‐β‐actin mouse mAb (8H10D10, #3700; Cell Signaling Technology, Danvers, Massachusetts)], anti‐BiP (rabbit Ab, 3177; Cell Signaling Technology), anti‐P4hb (rabbit Ab, ab137110; Abcam, Cambridge, UK), anti‐Calr (rabbit Ab, 2891; Cell Signaling Technology), anti‐GRP94 (rabbit Ab, 2104S; Cell Signaling Technology), anti‐GM130 (mouse Ab, 610 822; BD Biosciences, Franklin Lakes, New Jersey) and KDEL mouse antibody (2D6, MA5‐27581; Invitrogen, Carlsbad, California). The secondary antibodies used were goat anti‐rabbit IgG H&L Alexa Fluor 488 conjugated (ab150081; Abcam), goat anti‐mouse IgG H&L Alexa Fluor 647‐conjugated (ab150119; Abcam), HRP‐conjugated rabbit anti‐mouse IgG H&L (ab6728; Abcam) and goat anti‐rabbit IgG H&L (HRP) (ab6721; Abcam). The ER was stained using the ER‐ID Red assay kit, in accordance with the protocol of the manufacturer (Enzo Life Sciences, Farmingdale, New York).

Lysates were subjected to SDS-PAGE and transferred to nitrocellulose membranes. Total protein was quantified using REVERT stain (Li-Cor Biosciences). Primary antibodies were used according to manufacturer recommendations: anti-MUC16 [X75] (Novus Biologicals, NB6001468), anti-human cathepsin D (Cell Signaling, #2284), anti-mouse cathepsin D (Novus Biologicals, AF1029), anti-cathepsin B [D1C7Y] (Cell Signaling, #31718), anti-cathepsin L [33/2] (Santa Cruz, sc-32320), anti-LAMP1 [1D4B] (Developmental Studies Hybridoma Bank), anti-VDAC1 (Cell Signaling, #4661), anti-GOLGIN-97 (Cell Signaling, #13192), anti-CALR (Cell Signaling #12238), and anti-CATLASE (Cell Signaling, #14097). Secondary antibodies were used according to manufacturer recommendations.