Light cycler 2

To measure the mRNA levels of target genes, total RNA was isolated from cells by Qiagen lysis solution (Qiagen, Maryland, USA) and transcribed into cDNA by Go Script reverse transcription process (Promega) according to the manufacturer's instructions and as described previously^{67 (link)}. Quantitative real time-PCR was then performed using a Light Cycler 2.0 (Mannheim, Germany) using absolute QPCR SYBR green capillary mix system (Thermo Scientific, UK) at 95 °C for 15 min, with successive 40 cycles of 95 °C for 15 s, 56 °C for 30 s, and 72 °C for 45 s. The mRNA level of target gene was normalized to the value of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The sequences of the primers used for the PCR amplification are listed in Table 1.Table 1

Sequences of primers used in quantitative PCR.

Target gene	Primer	Nucleotide sequence
p62	F	5′-GCTCAGGAGGAGACGATGAC
	R	3′- AGAAACCCAAGGACAGCATC
TNF-α	F	5′-CCCTCACACTCAGATCATCTTCT-3′
	R	5′-GCTACGACGTGGGCTACAG-3′
GAPDH	F	5′-ACCACAGTCCATGCCATCAC-3′
	R	5′-TCCACCACCCTGTTGCTGTA-3′
IL-1β	F	5′-GCCTCGTGCTGTCGGACCCATAT
	R	3′-TCCTTTGAGGCCCAAGGCCACA
IL-6	F	5′-ACAACCACGGCCTTCCCTACTT
	R	3′-CACGATTTCCCAGAGAACATGTG
IFN-β	F	5′-AACTCCACCAGCAGACAGTG
	R	3′-TGAGGACATCTCCCACGTCA
Nrf2	F	5′-CTCGCTGGAAAAAGAAGTG
	R	3′-CCGTCCAGGAGTTCAGAGA
p21	F	5′-AATACCGTGGGTGTCAAAGC
	R	3′-GTGTGAGGACTCGGGACAAT
T-Cad	F	5′-TGCTGAAGACATGGCAGAAC
	R	3′-GGCTGACTCTGGTTCTCTGG

For the measurement of mRNA levels of target genes, total RNAs were isolated by Qiagen lysis solution (Qiagen, Maryland, USA) according to the manufacturer's instruction. Isolated total RNA was reverse transcribed into cDNA using Go Script reverse transcription system (Promega). Quantitative Real time-PCR was then performed using Light Cycler 2.0 (Mannheim, Germany) with the use of absolute QPCR SYBR green capillary mix AB gene system (Thermoscientific, UK) at 95°C for 15 s, 56°C for 30 s, and 72°C for 45 s. The amount of target mRNA was analyzed via comparative threshold (Ct) method after normalizing target mRNA Ct values to that for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ΔCt). The primer sequences used for amplification of target genes are listed in Table 1.