Mg132 s1748

The empty vectors pCMV-HA and pCMV-Myc were provided by the Biomedical Research Center of Northwest Minzu University. The recombinant plasmids Myc-HSP27, Myc-OAS1, Flag-MDA5, HA-VP1, HA-VP2, HA-2A, HA-2C, HA-3A, Myc-VP3, and Myc-3C were all constructed in the lab. The proteasome inhibitor MG132 (S1748) and caspase inhibitor Z-VAD-FMK (C1202) were obtained from Beyotime (Shanghai, China). The autolysosome inhibitor chloroquine (tlrl-chq) and poly (I:C) (tlrl-pic) were obtained from InvivoGen (San Diego, CA, United States). The small interfering RNA (siRNAs) an analog of double-stranded RNA (dsRNA) targeting homo HSP27 were designed and synthesized by Guangzhou Ribo Biotechnology (China). The siRNA sequences are listed in Table 1. A549 cells in 6-well plates were transfected with siRNAs or various plasmids using Lipofectamine™ 2000 according to the manufacturer’s instructions.

Rabbit polyclonal antibody (pAb) against Myc (R1208-1), GAPDH (glyceraldehyde-3-phosphate dehydrogenase; R1210-1), His (0812–1) and mouse mAbs against GST (M0807-1) were purchased from Huaan Biological Technology (Hangzhou, China). Mouse anti-Myc (05–419), anti-Flag (F1804) and anti-HA (H3663) mAbs for IP as well as chloroquine phosphate (CQ; 1118000) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit mAbs against TRIM21 (12108-1-AP) was purchased from Proteintech Group, Inc. (Manchester, UK). Mouse mAbs to PB2, NP, M1 and M2 were produced by our laboratory [62 (link)]. Horseradish peroxidase (HRP)-labeled goat anti-mouse or anti-rabbit IgG was purchased from KPL (Milford, MA, USA). Anti-mouse IgG HCS HRP-linked antibody and anti-Rabbit IgG LCS HRP-linked antibody (A25112 and A24022) were purchased from ABBkine Scientific (Wuhan, China). ChamQ Universal SYBR qPCR Master Mix (Q711-02) was purchased from Vazyme (Nanjing, China). NP-40 cell lysis buffer (P0013F) and MG132 (S1748) were purchased from Beyotime (Shanghai, China). MG132 was solubilized in Dimethyl sulfoxide (DMSO) to a stock of 25μM. PMSF (P0100) was purchased from Solarbio Biological Technology (Beijing, China). Protein A/G plus agarose (sc-2003) was purchased from Santa Cruz Biotechnology. GST resin (16100) was purchased from Thermo Fisher (Waltham, MA, USA).

Sangon Biotech (Shanghai, China) provided the following antibodies: rabbit polyclonal antibody against the FLAG tag (D191041), rabbit polyclonal antibody against the HA tag (D110004), rabbit polyclonal antibody against cGAS (D163570), HRP-conjugated Goat Anti-Rabbit IgG (D110058) and HRP-conjugated Goat Anti-Mouse IgG (D110087). The Polyclonal antibody for IRF3 (11312-1-AP) was acquired from Proteintech in Wuhan, China. The antibodies for phospho-IRF3 (Ser386) (37829 S), STING (13647 S), TBK1 (3013 S), and Myc-Tag (9B11) Mouse mAb (2276 S) were purchased from Cell Signaling Technology. The GAPDH Mouse Monoclonal Antibody (AF5009) and the β-actin Mouse Monoclonal Antibody (AA128) were purchased from Beyotime Biotechnology (Shanghai, China).
TransStart® Top Green qPCR SuperMix (+ Dye II) was bought from Transgen (Beijing, China). Invitrogen was the supplier of Lipofectamine 3000. The proteasome inhibitor MG132 (S1748) and the apoptosis inhibitor Z-VAD-FMK (C1202) were bought from Beyotime. InvivoGen provided the lysosomal pathway inhibitor chloroquine (CQ) (tlrl-chq), 2’3’-cGAMP (tlrl-nacga23-02), and poly (dA: dT) (tlrl-patc).