Journal: Neurotoxicity research

Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity

doi: 10.1007/s12640-020-00272-3

Figure Lengend Snippet: Chemicals used and abbreviations

Article Snippet: Chemicals Chemicals included Ferbam from TCI America, Portland, OR; ML385 and MS275 from Toris Biosciences, Minneapolis, MN; APTO235 (MedChemExpress, Princeton, NJ); and sankel (Muse Chem, Fairfield, NJ).

Techniques:

Journal: Neurotoxicity research

Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity

doi: 10.1007/s12640-020-00272-3

Figure Lengend Snippet: MT1G mRNA expression responses to chelated and unchelated metals. a dLUHMES cells were treated for 6 h with toxicants MHG, CuCl2, DDC, or ziram; and with vehicle, white bars; APTO-253 5 μM, speckled bars; or ML385 5 μM, dark hatched bars. MT1G expression was measured by qRT-PCR. Treatments were MHG 2 μM; CuCl2 5 μM; DDC 5 μM; NiDDC2 5 μM; ziram 5 μM; b undifferentiated LUHMES cells subjected to the same treatments

Article Snippet: Chemicals Chemicals included Ferbam from TCI America, Portland, OR; ML385 and MS275 from Toris Biosciences, Minneapolis, MN; APTO235 (MedChemExpress, Princeton, NJ); and sankel (Muse Chem, Fairfield, NJ).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Microbiology

Article Title: Replication of Dengue Virus in K562-Megakaryocytes Induces Suppression in the Accumulation of Reactive Oxygen Species

doi: 10.3389/fmicb.2021.784070

Figure Lengend Snippet: DENV replication suppresses ROS in differentiating cells by upregulation of NFE2L2 activity. (A) K562 cells either uninfected or infected with DENV at an MOI of 1.0 were differentiated with 50 nM PMA in the absence or presence of 10 μM of RA839. At 3 or 6 days post-infection the cytosolic ROS was quantified by H 2 DCFDA staining as described above. (B) K562 cells infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of RA839. The supernatant was collected at 3 or 6 days post-infection and the infectious titer of secreted virus quantified by Focus-forming unit (FFU) assay. The Log 10 of the FFU/ml was calculated and plotted. (C) K562 cells either uninfected or infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of ML385. At 3 or 6 days post-infection the cytosolic ROS was quantified by H 2 DCFDA staining as described above. (D) K562 cells infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of ML385. The culture supernatant was collected at 3 or 6 days post-infection and the infectious titer of secreted virus quantified by Focus-forming unit (FFU) assay. The Log 10 of the FFU/ml was calculated and plotted. (A–D) The error bars represent standard deviation obtained from 3 independent experiments and the P -values of significance calculated by Student's t -test are indicated.

Article Snippet: Phorbol-12 Myristate-13 acetate (PMA, Sigma Aldrich), N-Acetyl Cysteine (NAC, Sigma Aldrich), NITD008 (Sigma Aldrich), RA839 (Tocris), ML385 and Sodium Butyrate (Sigma Aldrich) were diluted in recommended vehicles and stored as single use aliquots at −20°C.

Techniques: Activity Assay, Infection, Staining, Virus, Standard Deviation

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho on renal function, inflammatory factors, oxidative stress and pathological kidney injury. (A,B) Levels of Scr (A) and BUN (B) in serum. n=6. (C,D) Levels of NGAL (C) and Kim-1 (D) in serum. n=6. (E,F) Levels of TNF-α (E) and IL-6 (F) in serum. n=6. (G-I) Levels of GSH-Px (G), T-SOD (H) and MDA (I) in serum. n=6. The results are expressed as the means ± standard deviation. *, P<0.05; **, P<0.01. CLP, cecal ligation and puncture; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein; Scr, serum creatinine; BUN, blood urea nitrogen; NGAL, neutrophil gelatinase-associated lipocalin; Kim-1, kidney injury molecule 1; TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; GSH-Px, glutathione peroxidase; SOD, superoxide dismutase; MDA, malondialdehyde.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Standard Deviation, Ligation, Recombinant

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho on pathological kidney injury. (A) HE staining scanning of renal tissue. (B) Histopathological scores. n=5. (C) Morphological observation of the kidney under an electron microscope. The results are expressed as the means ± standard deviation. *, P<0.05; ***, P<0.001; ****, P<0.0001. HE, hematoxylin-eosin; CLP, cecal ligation and puncture; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein; M, mitochondrion; N, nucleus.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Staining, Microscopy, Standard Deviation, Ligation, Recombinant

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho treatment on ferroptosis in CLP-induced SA-AKI. (A) Representative bands of WB and (B) quantification of GPX4 and Nrf2 in mouse kidneys in the sham and CLP groups. n=3. (C) Representative bands of WB and (D) quantification of GPX4 and Nrf2 in mouse kidneys among the four groups (sham, CLP, CLP + Lip-1, and CLP + Klotho groups). n=3. (E) Representative bands of WB and (F) quantification of GPX4 and Nrf2 among the four groups (sham, CLP, CLP + Klotho, and CLP + Klotho + Nrf2-IN-1 groups). β-actin was used as an internal control. The results are expressed as the means ± standard deviation. *, P<0.05; **, P<0.01. CLP, cecal ligation and puncture; SA-AKI, sepsis-associated acute kidney injury; WB, western blot; Nrf2, nuclear factor erythroid-2 related factor 2; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Control, Standard Deviation, Ligation, Western Blot, Recombinant

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho treatment on ferroptosis in CLP-induced SA-AKI. (A) Representative IF images and (B) quantification of GPX4 (red) in mouse kidneys from each group. n=3. (C) IF representative images and (D) quantification of Nrf2 (green) from mice in each group. n=3. (E) Levels of iron in tissue. n=6. The results are expressed as the means ± standard deviation. *, P<0.05; **, P<0.01. CLP, cecal ligation and puncture; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein; DAPI, 4,6-diamino-2-phenyl indole; Nrf2, nuclear factor erythroid-2 related factor 2; SA-AKI, sepsis-associated acute kidney injury; IF, immunofluorescence; ns, no significance.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Standard Deviation, Ligation, Recombinant, Immunofluorescence

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Klotho treatment increases the levels of Klotho in kidney tissue. (A) Representative bands and (B) quantification of Klotho in mouse kidneys in the sham and CLP groups. n=3. β-actin was used as an internal control. (C) Representative bands and (D) quantification of Klotho in mouse kidneys among the four groups (sham, CLP, CLP + Lip-1, and CLP + Klotho groups). n=3. (E) Representative bands and (F) quantification of Klotho among the four groups (sham, CLP, CLP + Klotho, and CLP + Klotho + Nrf2-IN-1 groups). n=3. β-actin was used as an internal control. (G) IHC staining and (H) quantification of Klotho. n=3. (I) Levels of Klotho in serum. n=6. The results are expressed as the means ± standard deviation. *, P<0.05; **, P<0.01. Klotho, recombinant human Klotho protein; CLP, cecal ligation and puncture; Lip-1, liprostatin-1; Nrf2, nuclear factor erythroid-2 related factor 2; IHC, immunohistochemical; ns, no significance.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Control, Immunohistochemistry, Standard Deviation, Recombinant, Ligation, Immunohistochemical staining

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho on LPS-induced HK2 cells. (A) Viability of HK2 cells in each group. n=3. (B1,B2) Levels of ROS in HK2 cells. n=3. (C) Representative bands and (D) quantification of GPX4 and Nrf2 in HK2 cells in the control and LPS groups. n=3. (E) Representative bands and (F) quantification of GPX4 and Nrf2 in HK2 cells in the control, LPS, LPS + Lip-1, and LPS + Klotho groups. n=3. (G) Representative bands and (H) quantification of GPX4 and Nrf2 in HK2 cells in the control, LPS, LPS + Klotho, and LPS + Klotho + Nrf2-IN-1 groups. n=3. β-actin was used as an internal control. *, P<0.05; **, P<0.01. LPS, lipopolysaccharide; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein; Nrf2, nuclear factor erythroid-2 related factor 2; ROS, reactive oxygen species; IF, immunofluorescence.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Control, Recombinant, Immunofluorescence

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Effect of Klotho on LPS-induced HK2 cells. (A) Representative IF images and (B) quantification of GPX4 (red) in HK2 cells in each group. n=3. (C) Representative IF images and (D) quantification of Nrf2 (red) in HK2 cells in each group. n=3. The results are expressed as the means ± standard deviation. ns, no significance; *, P<0.05; **, P<0.01. LPS, lipopolysaccharide; Lip-1, liprostatin-1; Klotho, recombinant human Klotho protein; Nrf2, nuclear factor erythroid-2 related factor 2; ROS, reactive oxygen species; IF, immunofluorescence; ns, no significance.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Standard Deviation, Recombinant, Immunofluorescence

Journal: Translational Andrology and Urology

Article Title: Klotho activation of Nrf2 inhibits the ferroptosis signaling pathway to ameliorate sepsis-associated acute kidney injury

doi: 10.21037/tau-23-573

Figure Lengend Snippet: Klotho treatment increases the levels of Klotho in HK2 cells. (A) Representative bands and (B) quantification of Klotho in HK2 cells in the control and LPS groups. n=3. (C) Representative bands and (D) quantification of Klotho in HK2 cells in the control, LPS, LPS + Lip-1, and LPS + Klotho groups. n=3. (E) Representative bands and (F) quantification of Klotho in HK2 cells in the control, LPS, LPS + Klotho, and LPS + Klotho + Nrf2-IN-1 groups. n=3. β-actin was used as an internal control. (G) Representative IF images and (H) quantification of Klotho (red) in HK2 cells in each group. n=3. The results are expressed as the means ± standard deviation. ns, no significance; *, P<0.05; **, P<0.01. Klotho, recombinant human Klotho protein; LPS, lipopolysaccharide; CLP, cecal ligation and puncture; Lip-1, liprostatin-1; Nrf2, nuclear factor erythroid-2 related factor 2; IF, immunofluorescence; ns, no significance.

Article Snippet: C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).

Techniques: Control, Standard Deviation, Recombinant, Ligation, Immunofluorescence

Journal: BMC Gastroenterology

Article Title: Emodin protects against severe acute pancreatitis-associated acute lung injury by activating Nrf2/HO-1/GPX4 signal and inhibiting ferroptosis in vivo and in vitro

doi: 10.1186/s12876-025-03660-1

Figure Lengend Snippet: Analysis of drug concentration of EMO and ML385 in A549 alveolar epithelial cells. ( A - B ) Cellular activity was assessed in response to different EMO concentrations using the CCK8 technique. ( C ) CCK8 detection of drug toxicity of ML385 on A549 cells. ( D ) The amount of Nrf2 mRNA expression in cells stimulated by varying ML385 concentration gradients was detected by PCR. *** p < 0.001

Article Snippet: ML385 was supplied by ApexBio Technology (USA).

Techniques: Concentration Assay, Activity Assay, Expressing