Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Protective effects of Rosavin on bleomycin-induced pulmonary fibrosis via suppressing fibrotic and inflammatory signaling pathways in mice.

doi: 10.1016/j.biopha.2019.108870

Figure Lengend Snippet: Fig. 8. The effect of Rosavin on the expression of TGF-β1, α-SMA, Nrf2 and NF-κB p65 of BIM-induced PF in mice. Values are the mean ± SD (n = 3); nsP > 0.05, ##P < 0.01 vs. control group. *P < 0.05, **P < 0.01 vs. Bleomycin control group.

Article Snippet: Rabbit anti-TGF-β1 antibody (21898-1-AP) was purchased from Proteintech (Wuhan, China), and mouse anti-α-SMA antibody (BM0002), rabbit anti-NF-Kb p65 antibody, rabbit anti-Nrf2 antibody were all obtained from Wuhan Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Control

Journal: Journal of Animal Science and Biotechnology

Article Title: Pyrroloquinoline quinone regulates the redox status in vitro and in vivo of weaned pigs via the Nrf2/HO-1 pathway

doi: 10.1186/s40104-021-00595-x

Figure Lengend Snippet: Effects of PQQ on proteins expression of Nrf2 and HO-1 in cells cultured with H 2 O 2 . a , Immunofluorescence staining of cytoplasmic and nuclear Nrf2 in IPEC-J2 cells. The scale bar represents 50 μm. b , Western blot analysis of nuclear Nrf2, cytoplasmic Nrf2, and HO-1. Densitometric values were normalized to those of PCNA or β-actin, as appropriate; c-e , Statistical analysis of the data in B. CTRL, cells cultured with basal medium; H 2 O 2 , cells cultured with 200 μmol/L H 2 O 2 for 2 h; PQQ, cells cultured with 10 nmol/L PQQ for 6 h and then incubated with both 200 μmol/L H 2 O 2 and 10 nmol/L PQQ for another 2 h. n = 3

Article Snippet: In brief, after 30 min of fixation with 4% paraformaldehyde solution, cells were permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% BSA for 1 h. Cells were incubated with a rabbit anti-Nrf2 antibody overnight at 4 °C and were then incubated with an Alexa Fluor 594-conjugated secondary antibody (ZF-0513, ZSGB-BIO) for 1 h. Subsequently, cell nuclei were stained with DAPI (Alexa Fluor® 555; ab150078; Abcam) for 10 min and imaged immediately using an Olympus fluorescence microscope (Tokyo, Japan).

Techniques: Expressing, Cell Culture, Immunofluorescence, Staining, Western Blot, Incubation

Journal: Journal of Animal Science and Biotechnology

Article Title: Pyrroloquinoline quinone regulates the redox status in vitro and in vivo of weaned pigs via the Nrf2/HO-1 pathway

doi: 10.1186/s40104-021-00595-x

Figure Lengend Snippet: Effects of PQQ on levels of Nrf2 and HO-1 in cells treated with siRNA. a Nrf2 knockdown efficiency in cells transfected with three different sequences of Nrf2 siRNA or with NC siRNA. b and c mRNA expression levels of Nrf2 and HO-1 in IPEC-J2 cells transfected with or without Nrf2 siRNA-2. d Western blot analysis of Nrf2 and HO-1 in cells transfected with or without Nrf2 siRNA-2. e Statistical analysis of the data in D. CTRL, cells cultured with basal medium; H 2 O 2 , cells cultured with 200 μmol/L H 2 O 2 for 2 h; PQQ, cells cultured with 10 nmol/L PQQ for 6 h and then incubated with both H 2 O 2 (200 μmol/L) and PQQ (10 nmol/L) for another 2 h. NC siRNA, cells transfected with NC siRNA and then pretreated with PQQ for 6 h prior to incubation with both PQQ and H 2 O 2 for another 2 h. Nrf2 siRNA, cells transfected with Nrf2 siRNA-2, pretreated with PQQ for 6 h, and then incubated with both PQQ and H 2 O 2 for another 2 h. * denotes a significant difference ( P < 0.05) with respect to the CTRL group. # denotes a significant difference ( P < 0.05) with respect to the H 2 O 2 group. n = 3

Article Snippet: In brief, after 30 min of fixation with 4% paraformaldehyde solution, cells were permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% BSA for 1 h. Cells were incubated with a rabbit anti-Nrf2 antibody overnight at 4 °C and were then incubated with an Alexa Fluor 594-conjugated secondary antibody (ZF-0513, ZSGB-BIO) for 1 h. Subsequently, cell nuclei were stained with DAPI (Alexa Fluor® 555; ab150078; Abcam) for 10 min and imaged immediately using an Olympus fluorescence microscope (Tokyo, Japan).

Techniques: Knockdown, Transfection, Expressing, Western Blot, Cell Culture, Incubation

Journal: Journal of Animal Science and Biotechnology

Article Title: Pyrroloquinoline quinone regulates the redox status in vitro and in vivo of weaned pigs via the Nrf2/HO-1 pathway

doi: 10.1186/s40104-021-00595-x

Figure Lengend Snippet: Effects of PQQ on levels of ROS and ROS-regulated proteins in cells treated with siRNA. a Level of ROS in IPEC-J2 cells treated with or without Nrf2 siRNA-2. b-c Ratio of Bcl-2/Bax mRNA levels and mRNA expression levels of Caspase-3 in IPEC-J2 cells treated with or without Nrf2 siRNA-2. d Western blot analysis of cytoplasmic Bax, Bcl-2 and Caspase-3. Densitometric values were normalized to those of β-actin. e Statistical analysis of the data in D. f Statistical analysis of the Bcl-2/Bax protein ratio. CTRL, cells cultured with basal medium; H 2 O 2 , cells cultured with 200 μmol/L H 2 O 2 for 2 h; PQQ, cells cultured with 10 nmol/L PQQ for 6 h and then incubated with both H 2 O 2 (200 μmol/L) and PQQ (10 nmol/L) for another 2 h. NC siRNA, cells transfected with NC siRNA and then pretreated with PQQ for 6 h prior to incubation with both PQQ and H 2 O 2 for another 2 h. Nrf2 siRNA, cells transfected with Nrf2 siRNA-2, pretreated with PQQ for 6 h, and then incubated with both PQQ and H 2 O 2 for another 2 h. n = 3

Article Snippet: In brief, after 30 min of fixation with 4% paraformaldehyde solution, cells were permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% BSA for 1 h. Cells were incubated with a rabbit anti-Nrf2 antibody overnight at 4 °C and were then incubated with an Alexa Fluor 594-conjugated secondary antibody (ZF-0513, ZSGB-BIO) for 1 h. Subsequently, cell nuclei were stained with DAPI (Alexa Fluor® 555; ab150078; Abcam) for 10 min and imaged immediately using an Olympus fluorescence microscope (Tokyo, Japan).

Techniques: Expressing, Western Blot, Cell Culture, Incubation, Transfection

Journal: Journal of Animal Science and Biotechnology

Article Title: Pyrroloquinoline quinone regulates the redox status in vitro and in vivo of weaned pigs via the Nrf2/HO-1 pathway

doi: 10.1186/s40104-021-00595-x

Figure Lengend Snippet: Effects of PQQ on tight junctions expression and cell viability in cells treated with siRNA. a Western blot analysis of ZO-1, ZO-2, Occludin and Claudin-1 in IPEC-J2 cells treated with or without Nrf2 siRNA-2. The densitometric values were normalized to those of β-actin. n = 3. b Viability of IPEC-J2 cells treated with or without Nrf2 siRNA-2 and incubated in the presence or absence of H 2 O 2 or PQQ. CTRL, cells cultured with basal medium; H 2 O 2 , cells cultured with 200 μmol/L H 2 O 2 for 2 h; PQQ, cells cultured with 10 nmol/L PQQ for 6 h and then incubated with both H 2 O 2 (200 μmol/L) and PQQ (10 nmol/L) for another 2 h. NC siRNA, cells transfected with NC siRNA and then pretreated with PQQ for 6 h prior to incubation with both PQQ and H 2 O 2 for another 2 h. Nrf2 siRNA, cells transfected with Nrf2 siRNA-2, pretreated with PQQ for 6 h, and then incubated with both PQQ and H 2 O 2 for another 2 h. n = 6. * denotes a significant difference ( P < 0.05) with respect to the CTRL group. # denotes a significant difference ( P < 0.05) with respect to the H 2 O 2 group

Article Snippet: In brief, after 30 min of fixation with 4% paraformaldehyde solution, cells were permeabilized with 0.2% Triton X-100 for 10 min and blocked with 5% BSA for 1 h. Cells were incubated with a rabbit anti-Nrf2 antibody overnight at 4 °C and were then incubated with an Alexa Fluor 594-conjugated secondary antibody (ZF-0513, ZSGB-BIO) for 1 h. Subsequently, cell nuclei were stained with DAPI (Alexa Fluor® 555; ab150078; Abcam) for 10 min and imaged immediately using an Olympus fluorescence microscope (Tokyo, Japan).

Techniques: Expressing, Western Blot, Incubation, Cell Culture, Transfection