Western blot analysis and Co-immunoprecipitation (co-IP) assay
Western blot and Co-IP were performed as described previously (L. Wang
et al., 2018). The primary antibodies for IRE1α (ab37073, 1:1000),
p-IRE1α (ab48187, 1:1000), PERK (ab79483, 1:1000), eIf2α (ab264254,
1:1000), ATF4 (ab23760, 1:1000), p38 (ab170099, 1:1000), p-p38
(ab178867, 1:1000) and p-Akt (ab38449, 1:800) were purchased from Abcam.
Antibodies against ATF6α (p50) (#65880, 1:1000), p-PERK (#3179,
1:1000) and p-eIf2α (#3398, 1:1000) were purchased from Cell Signaling.
Antibodies about CREB3L2 (14514-1-AP, 1:800), Akt (10176-2-AP, 1:2000),
GRP78 (11587-1-AP, 1:2000), GFP tag (50430-2-AP, 1:1500),
CHOP(15204-1-AP, 1:500), SREBP1 (14088-1-AP, 1:1000) were purchased from
Proteintech. β-Actin (AF5003, 1:1000) was purchased from Beyotime. was
purchased from Santa Cruz. Samples were subjected to standard SDS-PAGE,
and the resulting bands were transferred onto polyvinylidene difluoride
membrane for visualizing specific proteins. The ECL Plus detection
system (Thermo Scientific, USA) was used for detection.