Article Figures & Data
Figures
- Fig. 1.
Knockdown of lncRNAs HNF1α-AS1 and HNF4α-AS1 in HepaRG cells. HepaRG cells during a growth period were stably transfected with shNC, shHNF1α-AS1, or shHNF4α-AS1. (A) Relative expression of HNF1α-AS1 in HepaRG cells transfected with shNC or shHNF1α-AS1. (B) Relative expression of HNF4α-AS1 in HepaRG cells transfected with shNC or shHNF4α-AS1. Relative expression of HNF1α-AS1 and HNF4α-AS1 was measured by RT-PCR. The changes of relative mRNA expression compared with the shNC controls were calculated using the 2−∆∆t method after normalization with GAPDH. Data are shown as means ± S.D. (n = 3) and analyzed by two-tailed unpaired Student’s t test. **P < 0.01 and ***P < 0.001 vs. shNC controls.
- Fig. 2.
Changes of APAP-induced cytotoxicity by knockdown of HNF1α-AS1 and HNF4α-AS1 in HepaRG cells. (A) Cell viability evaluated by the MTT assay after APAP treatment at concentrations of 0, 10, 30, and 100 mM. (B) Cell damage assessed by the LDH release assay after APAP treatment. Data are shown as means ± S.D. (n = 3) and analyzed by a two-way ANOVA followed by Tukey’s test. *P < 0.05; **P < 0.01; ***P < 0.001 vs. shNC controls receiving different concentrations of APAP.
- Fig. 3.
Changes of APAP-induced cytotoxicity viewed by DHR123 staining in HepaRG cells with knockdown of lncRNAs HNF1α-AS1 and HNF4α-AS1. (A) Quantification of fluorescence intensity in different group of cells after treatment with 10 mM APAP. (B) The representative images are shown with green fluorescence in hepatocyte liver cells. Scale bar, 200 µM. Data are shown as means ± S.D. and analyzed by a one-way ANOVA analysis followed by Dunnett’s test. *P < 0.05 and **P < 0.01 vs. the shNC controls receiving 10 mM APAP.
- Fig. 4.
Metabolic pathways of APAP in liver by phase I and II enzymes.
- Fig. 5.
Impact on expression of APAP-metabolizing phase I enzymes in HepaRG cells with knockdown of HNF1α-AS1 and HNF4α-AS1. (A) Relative mRNA expression of CYP1A2, 2E1, and 3A4 was measured by RT-PCR. The changes of relative mRNA expression compared with the shNC controls were calculated using the 2−∆∆t method after normalization with GAPDH. Data are shown as means ± S.D. (n = 3) and analyzed by a one-way ANOVA analysis followed by Dunnett’s test. Three separate one-way ANOVAs were run. *P < 0.05; **P < 0.01; ***P < 0.001 vs. the shNC controls. (B) Protein expression of CYP1A2 and 2E1 was determined by Western blots. GAPDH was used as an internal control.
- Fig. 6.
Impact on expression of APAP-metabolizing phase II enzymes in HepaRG cells with knockdown of HNF1α-AS1 and HNF4α-AS1. (A) Relative mRNA expression of SULT1A1, UGT1A1, and UGT1A9 was measured by RT-PCR. (B) Relative mRNA expression of GSTP1 and GSTT1 was measured by RT-PCR. The changes of relative mRNA expression compared with the shNC controls were calculated using the 2−∆∆t method after normalization with GAPDH. Data are shown as means ± S.D. (n = 3) and analyzed by a one-way ANOVA analysis followed by Dunnett’s test. Three ANOVAs for the top panel and two for the bottom panel. *P < 0.05and **P < 0.01 vs. shNC controls.
- Fig. 7.
Mechanistic summary of HNF1α-AS1 and HNF4α-AS1 in susceptibility of AILI.
Additional Files
Data Supplement
- Supplemental Table -
Supplementary Table 1 - List of primers used for real-time PCR.
- Supplemental Table -
