Journal: Frontiers in Immunology

Article Title: Hyperglycemia Aggravates Hepatic Ischemia and Reperfusion Injury by Inhibiting Liver-Resident Macrophage M2 Polarization via C/EBP Homologous Protein-Mediated Endoplasmic Reticulum Stress

doi: 10.3389/fimmu.2017.01299

Figure Lengend Snippet: Hyperglycemia enhances endoplasmic reticulum stress in livers post-IR. Diabetic [streptozotocin (STZ)] and control (CON) mice were prepared and liver partial warm ischemia and reperfusion (IR) or sham procedure was performed. Six hours post-reperfusion, liver tissues were collected, and c-ATF6, ATF4, C/EBP homologous protein (CHOP), s-XBP1, and β-actin protein levels were analyzed by Western blotting. Representative of three experiments (A) . Relative density ratios of target proteins in different groups to the control group (CON-Sham) were calculated [ (B) , n = 3/group] (* p < 0.05).

Article Snippet: Primary antibodies against cleaved-ATF6 (c-ATF6, Novus, Littleton, CO, USA), ATF4 (Proteintech Group, Chicago, IL, USA), CHOP (Cell Signaling Technology, MA, USA), spliced XBP1 (s-XBP1, Abcam, Cambridge, MA, USA), and β-actin (Cell Signaling Technology, MA, USA) were used and incubated overnight at 4°C.

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Hyperglycemia Aggravates Hepatic Ischemia and Reperfusion Injury by Inhibiting Liver-Resident Macrophage M2 Polarization via C/EBP Homologous Protein-Mediated Endoplasmic Reticulum Stress

doi: 10.3389/fimmu.2017.01299

Figure Lengend Snippet: C/EBP homologous protein (CHOP) mediates hyperglycemic Kupffer cell (KC) pro-inflammatory activation in vitro . Diabetic [streptozotocin (STZ)] and control (CON) mice were prepared and liver partial warm ischemia and reperfusion (IR) or a sham procedure was performed. After 6 h of reperfusion, KCs were isolated and the intracellular levels of c-ATF6, ATF4, CHOP, s-XBP1, and β-actin protein were analyzed by Western blotting. Representative of three experiments (A) . Relative density ratios of target proteins in different groups to the control group (CON-Sham) were calculated [ (B) , n = 3/group]. Both CON and STZ mice were pretreated with CHOP siRNA (CHOP-siRNA) or its scramble control siRNA (SCR-siRNA) in vivo prior to IR using mannose-conjugated polymers as described in Section “ .” Liver IR was performed. Six hours post-reperfusion, KCs were isolated and the intracellular levels of c-ATF6, ATF4, CHOP, s-XBP1, and β-actin protein were analyzed by Western blotting. Representative of three experiments (C) . Relative density ratios of target proteins in different groups to the control group (CON–SCR-siRNA) were calculated [ (D) , n = 3/group]. Isolated KCs from IR-stressed livers of different groups were cultured for 6 h, and TNF-α, IL-6, and IL-10 protein levels in the culture supernatant were measured by ELISA [ (E) , n = 6/group] (* p < 0.05).

Article Snippet: Primary antibodies against cleaved-ATF6 (c-ATF6, Novus, Littleton, CO, USA), ATF4 (Proteintech Group, Chicago, IL, USA), CHOP (Cell Signaling Technology, MA, USA), spliced XBP1 (s-XBP1, Abcam, Cambridge, MA, USA), and β-actin (Cell Signaling Technology, MA, USA) were used and incubated overnight at 4°C.

Techniques: Activation Assay, In Vitro, Isolation, Western Blot, In Vivo, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: Gene primer sequences.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: Effects of GC on CP-induced ER stress. (A) Effect of GC on the morphological changes of ER in liver cells induced by CP. Red arrows show the morphology of the endoplasmic reticulum and ribosomes. (B, C) Effect of GC on CP-induced ER stress-related indicators. GRP78, ATF6, and p-IRE1α protein expression were detected by WB analysis. GRP78 and ATF6 gene expression were detected by qRT-PCR analysis. All data are presented as the mean ± SD. ## p < 0.01 compared with control group; * p < 0.05, ** p < 0.01 compared with CP group.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Control

Journal: Frontiers in Pharmacology

Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis

doi: 10.3389/fphar.2025.1557125

Figure Lengend Snippet: (A-C) MTT assay for determining cell viability. (D-E) GC reduced the expression of ER stress-related indicators. Expression levels of GRP78, ATF6, and p-IRE1α protein were tested by WB analysis. Expression levels of GRP78 and ATF6 mRNA were tested by qRT-PCR analysis.

Article Snippet: WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech).

Techniques: MTT Assay, Expressing, Quantitative RT-PCR

Journal: Cell & Bioscience

Article Title: Exogenous H 2 S restores ischemic post-conditioning-induced cardioprotection through inhibiting endoplasmic reticulum stress in the aged cardiomyocytes

doi: 10.1186/s13578-017-0196-9

Figure Lengend Snippet: Low dose of H 2 O 2 induced senescence in the H9C2 cells without causing apoptosis. a The AGEs content. The different concentrations (0, 5, 10, 30, 50, 80, 100 μM) H 2 O 2 was added to the H9C2 cells in the culture cluster for 2 h and subsequently cultured for 3 days. All data were from eight independent experiments. *p < 0.05, **p < 0.01 vs . control group. b The caspase-3 activity. Caspase-3 activity, measured as luminescence, in the H9C2 cells 3 days after treatment with H 2 O 2 (30, 50, 80, 100 μM) for 2 h. All data were from four independent experiments. **p < 0.01 vs . control group. c The number of SA β-gal-positive H9C2 cells. Phase-contrast images showing morphologic changes and stained cells. (SA-β-gal positive cells; Blue, magnification ×400). Scale bar = 50 μm. The aged H9C2 cells in at least five random fields were counted. **p < 0.01 vs . control group. d The expression of cyclin D1 and p21 Cip/WAF−1 . The intensity of each band was quantified by densitometry, and data were normalized to the GAPDH signal. All data were from four independent experiments. *p < 0.05 vs . control group

Article Snippet: The primary antibodies for anti-Cyclin D1, p21 Cip/WAF−1 , cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c ), GRP 78, CHOP, cleaved caspase-12, ATF 4, ATF 6, XBP-1 and GAPDH were from Proteintech (Wuhan, China).

Techniques: Cell Culture, Control, Activity Assay, Staining, Expressing

Journal: Cell & Bioscience

Article Title: Exogenous H 2 S restores ischemic post-conditioning-induced cardioprotection through inhibiting endoplasmic reticulum stress in the aged cardiomyocytes

doi: 10.1186/s13578-017-0196-9

Figure Lengend Snippet: The effect of exogenous H 2 S on the expression of apoptotic relative factors. a Cleaved caspase-9; b Cleaved caspase-3; c Cyt c ; d Bcl-2. The intensity of each band was quantified by densitometry, and data were normalized to the GAPDH signal. The expression levels in the control group were considered the basal levels, and the others are expressed as fold change from the control group. The fold change values represent the mean ± S.E.M. of four determinations. *p < 0.05, **p < 0.01 vs . control group; # p < 0.05 vs . H/R group; && p < 0.01 vs . PC group; $ p < 0.05 vs . H/R + NaHS group

Article Snippet: The primary antibodies for anti-Cyclin D1, p21 Cip/WAF−1 , cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c ), GRP 78, CHOP, cleaved caspase-12, ATF 4, ATF 6, XBP-1 and GAPDH were from Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Cell & Bioscience

Article Title: Exogenous H 2 S restores ischemic post-conditioning-induced cardioprotection through inhibiting endoplasmic reticulum stress in the aged cardiomyocytes

doi: 10.1186/s13578-017-0196-9

Figure Lengend Snippet: Exogenous H 2 S inhibited the expression of the relative proteins of ERS. a GRP 78; b CHOP; c Cleaved caspase-12; d XBP-1. The intensity of each band was quantified by densitometry, and data were normalized to the GAPDH signal. The expression levels in the control group were considered the basal levels, and the others are expressed as fold change from the control group. The fold change values represent the mean ± S.E.M. of four determinations. **p < 0.01 vs . control group; # p < 0.05, ## p < 0.01 vs . H/R group; && p < 0.01 vs . PC group; $ p < 0.05 vs . H/R + NaHS group

Article Snippet: The primary antibodies for anti-Cyclin D1, p21 Cip/WAF−1 , cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c ), GRP 78, CHOP, cleaved caspase-12, ATF 4, ATF 6, XBP-1 and GAPDH were from Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Cell & Bioscience

Article Title: Exogenous H 2 S restores ischemic post-conditioning-induced cardioprotection through inhibiting endoplasmic reticulum stress in the aged cardiomyocytes

doi: 10.1186/s13578-017-0196-9

Figure Lengend Snippet: Exogenous H 2 S inhibited PERK-eIF 2α-ATF 4 pathway. a PERK; b eIF 2α; c ATF 4; d ATF 6. The graphs represent the optical density of the bands of phosphorylated PERK and eIF 2α normalized with the expression of total PERK and eIF 2α, respectively. The graphs represent the optical density of the bands of ATF 4 and ATF 6 normalized with the expression of GAPDH signal. The expression levels in the control group were considered the basal levels, and the others are expressed as fold change from the control group. The fold change values represent the mean ± S.E.M. of three determinations. *p < 0.05, **p < 0.01 vs . control group; # p < 0.05, ## p < 0.01 vs . H/R group; && p < 0.01 vs . PC group; $ p < 0.05 vs . H/R + NaHS group

Article Snippet: The primary antibodies for anti-Cyclin D1, p21 Cip/WAF−1 , cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c ), GRP 78, CHOP, cleaved caspase-12, ATF 4, ATF 6, XBP-1 and GAPDH were from Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Cell & Bioscience

Article Title: Exogenous H 2 S restores ischemic post-conditioning-induced cardioprotection through inhibiting endoplasmic reticulum stress in the aged cardiomyocytes

doi: 10.1186/s13578-017-0196-9

Figure Lengend Snippet: Exogenous H 2 S down-regulated IRE 1α-XBP-1 pathway. a IRE 1α; b XBP-1. The graphs represent the optical density of the bands of phosphorylated IRE 1α normalized with the expression of total IRE 1α. The graphs represent the optical density of the bands of XBP-1 normalized with the expression of GAPDH signal. The expression levels in the control group were considered the basal levels, and the others are expressed as fold change from the control group. The fold change values represent the mean ± S.E.M. of three determinations. **p < 0.01 vs . control group; # p < 0.05, ## p < 0.01 vs . H/R group; && p < 0.01 vs . PC group; $ p < 0.05 vs . H/R + NaHS group

Article Snippet: The primary antibodies for anti-Cyclin D1, p21 Cip/WAF−1 , cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c ), GRP 78, CHOP, cleaved caspase-12, ATF 4, ATF 6, XBP-1 and GAPDH were from Proteintech (Wuhan, China).

Techniques: Expressing, Control