Journal: Cell reports

Article Title: Stress-induced perturbations in intracellular amino acids reprogram mRNA translation in osmoadaptation independently of the ISR

doi: 10.1016/j.celrep.2022.111092

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-S6 , Cell Signaling Technology , Cat# 2895; RRID: AB_2089254.

Techniques: Recombinant, Protease Inhibitor, Labeling, Transfection, Clone Assay, Staining, Multiplex Assay, Reverse Transcription, Footprinting, Subcloning, Plasmid Preparation, shRNA, Control, Software

Journal: Renal Failure

Article Title: Apelin-13 alleviates contrast-induced acute kidney injury by inhibiting endoplasmic reticulum stress

doi: 10.1080/0886022X.2023.2179852

Figure Lengend Snippet: Iohexol induces ER stress, oxidative stress and apoptosis in rat kidney tissues. The method for building the CI-AKI rat model has been described in our previous article. (A, B) Changes in the levels of Scr and BUN. (C) Renal GSH content. (D) Renal MDA activity. (E, F) Representative immunoblot analysis and semi-quantitative analysis of GRP78, CHOP, caspase-12 and Cleaved caspase-3, GAPDH was used as a loading control. Data are expressed as means ± SEMs. n = 4. * p < 0.05, ** p < 0.01, significantly different from control group.

Article Snippet: Anti-CHOP (15204-1-AP), anti-78-kDa glucose-regulated protein (GRP78) (66574-1-Ig), anti-Nuclear respiratory factor 2 (Nrf2) (16396-1-AP), anti-Keap1 (60027-1-Ig), anti-GAPDH (10494-1-AP) and anti-β-actin (20536-1-AP) were obtained from Proteintech (Chicago, USA).

Techniques: Activity Assay, Western Blot

Journal: Renal Failure

Article Title: Apelin-13 alleviates contrast-induced acute kidney injury by inhibiting endoplasmic reticulum stress

doi: 10.1080/0886022X.2023.2179852

Figure Lengend Snippet: Apelin-13 attenuated iohexol-induced ER stress in rat tubular cells. Rats were treated as described above. The expression of GRP78, p-PERK, PERK and CHOP in kidney tissues was detected by immunoblot analysis. Immunofluorescence was used to detect the expression of GRP78 and CHOP in kidney tissues. (A, B) Representative immunoblot analysis and semi-quantitative analysis of GRP78, p-PERK, PERK, CHOP in rat kidney tissues, GAPDH was used as a loading control. (C) Representative immunofluorescence images of the GRP78 and CHOP. * p < 0.05, ** p < 0.01, significantly different from control group; # p < 0.05, ## p < 0.01, significantly different from the CM group. All quantitative data are expressed as means ± SEMs, n = 3.

Article Snippet: Anti-CHOP (15204-1-AP), anti-78-kDa glucose-regulated protein (GRP78) (66574-1-Ig), anti-Nuclear respiratory factor 2 (Nrf2) (16396-1-AP), anti-Keap1 (60027-1-Ig), anti-GAPDH (10494-1-AP) and anti-β-actin (20536-1-AP) were obtained from Proteintech (Chicago, USA).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Renal Failure

Article Title: Apelin-13 alleviates contrast-induced acute kidney injury by inhibiting endoplasmic reticulum stress

doi: 10.1080/0886022X.2023.2179852

Figure Lengend Snippet: Apelin-13 attenuates iohexol-induced ER stress, oxidative stress and apoptosis in HK-2 cells. (A, B) HK-2 cells were incubated in medium containing 200 mg iodine/mL iohexol and / or interfered with different concentrations of apelin-13 (0, 0.1, 1, 10, 100, 1000 nM) for 6 h . HK-2 cells were incubated in the medium containing 200 mg iodine/mL iohexol for 6 h and / or given the intervention of 100 nM apelin-13 at indicated time points (0 h, 3 h, 6 h, 9 h, 12 h) prior to cell collection. The cells cultured in a normal medium without iohexol were used as a control. Cell viability was detected with CCK-8 assay. Cell viability of the control group was set to 100 %, and other groups were normalized to indicate cell viability changes with the control group ( n = 8). (C - H) HK 2 cells were incubated in medium containing 200 mg iodine/mL iohexol and apelin(10 nM, 1000 nM) for 6 h. Cells cultured in the normal medium without iohexol or apelin-13 were used as a control. (C) Cell GSH content. (D) Cell MDA activity. (E - H) Representative bands of p-PERK, CHOP, caspase-4, Cleaved caspase-3, Keap1, Nrf2 and semi-quantitative analysis of these protein expression. * p < 0.05, ** p < 0.01, significantly different from control group; # p < 0.05, ## p < 0.01, significantly different from the CM group. Data are expressed as means ± SEMs.

Article Snippet: Anti-CHOP (15204-1-AP), anti-78-kDa glucose-regulated protein (GRP78) (66574-1-Ig), anti-Nuclear respiratory factor 2 (Nrf2) (16396-1-AP), anti-Keap1 (60027-1-Ig), anti-GAPDH (10494-1-AP) and anti-β-actin (20536-1-AP) were obtained from Proteintech (Chicago, USA).

Techniques: Incubation, Cell Culture, CCK-8 Assay, Activity Assay, Expressing

Journal: Acta Veterinaria Scandinavica

Article Title: Effects of osteoprotegerin from transfection of pcDNA3.1(+)/chOPG on bioactivity of chicken osteoclasts

doi: 10.1186/1751-0147-53-21

Figure Lengend Snippet: Gel electrophoresis of chOPG. 1A : Gel electrophoresis of reverse transcription polymerase chain reaction (RT-PCR) product. Total RNA extracted from chicken embryo frontal bone was analyzed using RT-PCR with specific primers. About 1.2 kbp gene of chicken osteoprotegerin (chOPG) was amplified (lane 1); DL2000 marker (lane 2); 1B : Gel electrophoresis of pcDNA3.1 (+)/chOPG PCR product. chOPG fragment was inserted into the eucaryon expression vector pcDNA3.1 (+) between Nhe | and Xho |. Negative plasmid (lane 1) and positive plasmid (lane 2) were chosen using PCR; marker (lane 3); 1C : Gel electrophoresis of pcDNA3.1 (+)/chOPG double restriction enzyme assay. Positive plasmid (lane 3) was identified by Nhe |and Xho |double restriction digestion and showed pcDNA3.1 (+) and OPG (lane 2); marker (lane 1); 1D : Gel electrophoresis of RT-PCR analysis showing the expression of chOPG gene at 48 h. Amplification of chOPG using cDNA from lane 1 (control group) and lane 2 (pcDNA3.1 (+) transfected CEFs group) showing negative result. Amplification of chOPG using cDNA from lane 4 (pcDNA3.1 (+)/chOPG transfected chicken embryo fibroblasts group) showing about 1200 bp gene of chOPG. Lane 3: marker

Article Snippet: Detergent extraction with 3% Triton X-100 was performed for 10 min. Coverslips were saturated with PBS containing 5% bovine serum albumin (Wuhan Boster Biotechnology Company, China) for 1 h at room temperature with gentle rocking, processed with rabbit anti-chOPG polyclonal antibody (Nanjing Agricultural University) for 1 h at 37°C and followed by FITC-goat-anti-rabbit IgG (Wuhan Boster Biotechnology Company) for 1 h at 37°C and then stained by DAPI staining solution (Wuhan Boster Biotechnology Company).

Techniques: Nucleic Acid Electrophoresis, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Expressing, Plasmid Preparation, Enzymatic Assay, Control, Transfection

Journal: Acta Veterinaria Scandinavica

Article Title: Effects of osteoprotegerin from transfection of pcDNA3.1(+)/chOPG on bioactivity of chicken osteoclasts

doi: 10.1186/1751-0147-53-21

Figure Lengend Snippet: The expression of chOPG protein and effect on osteoclast morphology, apoptosis and resorption. 2A : Immunofluorescence assay for a possible chicken osteoprotegerin (chOPG) protein. Chicken embryo fibroblasts (CEFs) were grown on coverslips, fixed, and examined by indirect immunofluorescence. Cells were incubated with rabbit anti-chOPG serum. The secondary antibody was fluorescein-conjugated goat anti-rabbit immunoglobulin G (green). The nuclei of the corresponding cells were visualized by DAPI staining (blue). Fluorescence signals were analyzed by Fluoview microscopy (×200). Negative results are shown on card l (control group) and card 2 (pcDNA3.1 (+) transfected CEFs group), positive green fluorescence for CEFs are shown on card 3 (pcDNA3.1 (+)/chOPG transfected CEFs group). 2B : The morphology of osteoclasts was observed by inverted phase contrast microscope (×200). The adherent osteoclasts were cultured in Dulbecco's modified Eagle's medium (DMEM) containing supernatant of control group (l), pcDNA3.1 (+) transfected CEF group (2) and pcDNA3.1 (+)/chOPG transfected CEF group (3) for 5 d. 2C : Effect of the supernatant of three groups on the apoptosis of osteoclasts by flow cytometry. 2D : Toluidine blue staining of bone slices showing resorption lacunae (×200). The adherent osteoclasts were cultured in DMEM containing supernatant for 5 d in three groups.

Article Snippet: Detergent extraction with 3% Triton X-100 was performed for 10 min. Coverslips were saturated with PBS containing 5% bovine serum albumin (Wuhan Boster Biotechnology Company, China) for 1 h at room temperature with gentle rocking, processed with rabbit anti-chOPG polyclonal antibody (Nanjing Agricultural University) for 1 h at 37°C and followed by FITC-goat-anti-rabbit IgG (Wuhan Boster Biotechnology Company) for 1 h at 37°C and then stained by DAPI staining solution (Wuhan Boster Biotechnology Company).

Techniques: Expressing, Immunofluorescence, Incubation, Staining, Fluorescence, Microscopy, Control, Transfection, Cell Culture, Modification, Flow Cytometry

Journal: Acta Veterinaria Scandinavica

Article Title: Effects of osteoprotegerin from transfection of pcDNA3.1(+)/chOPG on bioactivity of chicken osteoclasts

doi: 10.1186/1751-0147-53-21

Figure Lengend Snippet: Effect of culture supernatant of chicken embryo fibroblasts on the quantity and area of osteoclast resorption lacunae in three groups.

Article Snippet: Detergent extraction with 3% Triton X-100 was performed for 10 min. Coverslips were saturated with PBS containing 5% bovine serum albumin (Wuhan Boster Biotechnology Company, China) for 1 h at room temperature with gentle rocking, processed with rabbit anti-chOPG polyclonal antibody (Nanjing Agricultural University) for 1 h at 37°C and followed by FITC-goat-anti-rabbit IgG (Wuhan Boster Biotechnology Company) for 1 h at 37°C and then stained by DAPI staining solution (Wuhan Boster Biotechnology Company).

Techniques: