Prime script rt master mix

Nucleospin RNA Kit (Takara, Dalian, China) was used to extract the total RNA from samples. cDNA was synthesized using Primescript RT Master Mix (Transgen, Beijing, China) obeying the manufacturer’s guidance. qPCR was launched using SYBR Premix Ex Taq II (Vazyme, Nanjing, China) and Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Foster City, USA) to measure HSV-1 virus DNA. The cycle condition was 95°C for 30s, followed by 40 two-step cycles (95°C for 5s, 60°C for 30s). The primers for the genes RPL5, HSV-1 ICP0, TK, VP16, mrpl5, il-6, il-1α, tnf-α, nlrp3, caspase 1 and il-1β are provided in Table 1 and the relative gene expression was determined based on deactivation with the housekeeping gene as described previously.30 (link)
Table 1

Primers Used for qPCR

No.	Gene	Forward Primer	Reverse Primer
1.	RPL5	GGAAGCACATCATGGGTCAGA	TACGCATCTTCATCTTCCTCCATT
2.	ICP0	GGCCCCCTTGTCAACAGA	GGGAGTCGCTGATCACTATGG
3.	VP16	GCGGGGCCGGGATTTACC	CTCGAAGTCGGCCATATCCA
4.	TK	AAGGTCGGCGGGATGAG	CGGCCGCGCGATAC
5.	il1b	CTTTCCCGTGGACCTTCCA	CTCGGAGCCTGTAGTGCAGTT
6.	tnf-a	ACAAGGCTGCCCCGACTAC	TGGGCTCATACCAGGGTTTG
7.	il6	ACCACTCCCAACAGACCTGTCT	CAGATTGTTTTCTGCAAGTGCAT
8.	il1a	GATGAAGCTCGTCAGGCAGAA	CCTCCCGACGAGTAGGCATA
9.	casp1	CTGGGACCCTCAAGTTTTGC	CCCTCGGAGAAAGATGTTGAAA
10.	m-rpl5	CCGCAGGCTTCTGAATAGGT	CCAGTTGTAGTTCGGGCAAGA
11.	nlrp3	CTGCGGACTGTCCCATCAAT	AGGTTGCAGAGCAGGTGCTT

DMEM and penicillin-streptomycin-glutamine (100×) were purchased from Gibco (Waltham, MA, USA). Fetal bovine serum (FBS) was purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China). Exosome-depleted FBS was purchased from Cellmax Cell Technology Co., Ltd. (Beijing, China). Lefty1 recombinant plasmid (Gene ID: 498299), negative plasmid and GAPDH primers, U6 primers, α-SMA primers, lefty1 primers were purchased from Genepharma Technology Co., Ltd (Shanghai, China). PKH26 was got from Sigma (St. Louis, MO, USA). TransExo™ serum/plasma exosome total RNA extraction kit and PrimeScript RT master mix were got from TransGen Biotech Co., Ltd (Beijing, China). Plasmocin™ prophylactic, dextran-AlexaFluor555, transferrin-AlexaFluor 555, cholera toxin subunit B-AlexaFluor 555 and DiR were purchased from Invivogen (San Diego, CA, USA). Hoechst33342 and lysotracker green were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Anti-CD9(ab92726), CD81(ab109201), TSG101 (ab125011), α-SMA(ab7817), collagen I(ab270993) and HRP coupling goat anti-rabbit IgG (ab205718), goat anti-rabbit IgG H&L (Alexa Fluor 488) (ab150077) were purchased from Abcam (Cambridge, UK). Anti-alix (T57215) was purchased from Abmart Medical Technology Co., Ltd. (Shanghai, China). Anti-CD63(AF5117), calnexin (AF5362) were purchased from Affinity Biosciences, Inc. (Cincinnati, OH, USA).