Journal: Frontiers in Pharmacology

Article Title: Role of albumin in regulating platelet function

doi: 10.3389/fphar.2026.1734694

Figure Lengend Snippet: Hypoproteinemia was found to promote platelet activation. (A–E) Wash platelet aggregation in WT, ADR and ADR Albumin mice induced by thrombin (A) , U46619 (B) , PAR4-AP (C) , collagen (D) and ADP (E) (n = 5 independent experimental animals). (F–H) Thrombin (F) , U46619 (G) and PAR4-AP (H) induced integrin αIIbβ3 activation in washed platelets from mice (n = 5 independent experimental animals). (I–K) Thrombin (I) , U46619 (J) and PAR4-AP (K) induced P-selectin release from washed platelets from mice (n = 5 independent experimental animals). (L–O) ATP release from washed platelets in mice was induced by thrombin (L) , U46619 (M) , PAR4-AP (N) and collagen (O) (n = 5 independent experimental animals). (P–R) Wash platelet spreading in mice induced by U46619 (P) , mean spreading area of individual platelets (Q) , and proportion of spreading platelets to total platelets in the field of view (R) (n = 3 independent experimental animals). Differences between groups were assessed by one-way ANOVA followed by Dunnett’s post hoc test. Statistics are presented as mean ± SEM. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: Rabbit antibodies against phospho-PKC substrate (2261), phospho-Akt (Ser473) (11E7), Akt (pan) (11E7), PKC alpha (2056), GAPDH (5174), cell lysate buffer (9803), and PAR4-AP (2328) were sourced from Cell Signaling Technology, Inc. (Beverly, MA, United States).

Techniques: Activation Assay

Journal: Korean Circulation Journal

Article Title: UPF1 Alleviates Myocardial Ischemia-Reperfusion Injury by Regulating SMURF2 -Mediated Ubiquitination Degradation of FOXA2

doi: 10.4070/kcj.2024.0190

Figure Lengend Snippet: DMEM = Dulbecco's Modified Eagle's Medium; FOXA2 = forkhead box A2; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; H/R = hypoxia/reoxygenation; IgG = immunoglobulin G; NC = negative control; oe = overexpression; PAR4 = protease-activated receptor 4; RT-qPCR = reverse transcription quantitative polymerase chain reaction. ** p<0.01, *** p<0.001.

Article Snippet: After 1 hour of blockade with 5% bovine serum albumin, membranes were incubated with antibodies overnight at 4˚C: UPF1 (#12040, Cell Signaling Technology, CST, Danvers, MA, USA), FOXA2 (#8186T, 1:1,000, CST), PAR4 (#2328, 1:1,000, CST), Bax (ab32503, 1:1,000, Abcam, Cambridge, MA, USA), Cleaved caspase-3 (ab184787, 1:1,000, Abcam), SYVN1 (13473-1-AP, 1:1,000, Proteintech, Rosemont, IL, USA), SMURF2 (18038-1-AP, 1:1,000, Proteintech), WW domain containing E3 ubiquitin protein ligase 2 (WWP2; 12197-1-AP, 1:1,000, Proteintech).

Techniques: Modification, Negative Control, Over Expression, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Korean Circulation Journal

Article Title: UPF1 Alleviates Myocardial Ischemia-Reperfusion Injury by Regulating SMURF2 -Mediated Ubiquitination Degradation of FOXA2

doi: 10.4070/kcj.2024.0190

Figure Lengend Snippet: GAPDH = glyceraldehyde 3-phosphate dehydrogenase; FOXA2 = forkhead box A2; I/R = ischemia/reperfusion; NC = negative control; oe = overexpression; PAR4 = protease-activated receptor 4; SMURF2 = SMAD-specific E3 ubiquitin ligase 2; TTC = tetrazolium chloride; UPF1 = up-frameshift 1. *** p<0.001.

Article Snippet: After 1 hour of blockade with 5% bovine serum albumin, membranes were incubated with antibodies overnight at 4˚C: UPF1 (#12040, Cell Signaling Technology, CST, Danvers, MA, USA), FOXA2 (#8186T, 1:1,000, CST), PAR4 (#2328, 1:1,000, CST), Bax (ab32503, 1:1,000, Abcam, Cambridge, MA, USA), Cleaved caspase-3 (ab184787, 1:1,000, Abcam), SYVN1 (13473-1-AP, 1:1,000, Proteintech, Rosemont, IL, USA), SMURF2 (18038-1-AP, 1:1,000, Proteintech), WW domain containing E3 ubiquitin protein ligase 2 (WWP2; 12197-1-AP, 1:1,000, Proteintech).

Techniques: Negative Control, Over Expression

Journal: Korean Circulation Journal

Article Title: UPF1 Alleviates Myocardial Ischemia-Reperfusion Injury by Regulating SMURF2 -Mediated Ubiquitination Degradation of FOXA2

doi: 10.4070/kcj.2024.0190

Figure Lengend Snippet: FOXA2 = forkhead box A2; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; H/R = hypoxia/reoxygenation; NC = negative control; oe = overexpression; PAR4 = protease-activated receptor 4; SMURF2 = SMAD-specific E3 ubiquitin ligase 2; UPF1 = up-frameshift 1. * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: After 1 hour of blockade with 5% bovine serum albumin, membranes were incubated with antibodies overnight at 4˚C: UPF1 (#12040, Cell Signaling Technology, CST, Danvers, MA, USA), FOXA2 (#8186T, 1:1,000, CST), PAR4 (#2328, 1:1,000, CST), Bax (ab32503, 1:1,000, Abcam, Cambridge, MA, USA), Cleaved caspase-3 (ab184787, 1:1,000, Abcam), SYVN1 (13473-1-AP, 1:1,000, Proteintech, Rosemont, IL, USA), SMURF2 (18038-1-AP, 1:1,000, Proteintech), WW domain containing E3 ubiquitin protein ligase 2 (WWP2; 12197-1-AP, 1:1,000, Proteintech).

Techniques: Negative Control, Over Expression

Journal: Antioxidants

Article Title: Nrf2 Activation Does Not Protect from Aldosterone-Induced Kidney Damage in Mice

doi: 10.3390/antiox12030777

Figure Lengend Snippet: Abundance of Nrf2 in Aldo-infused WT and Nrf2ꜛ mice and control mice after 28 days of treatment. Representative pictures of the Western blots of the expression of total Nrf2 (95–110 kDa) in ( a ) nuclear and ( b ) cytosolic extracts of kidneys of mice as well as the quantification of band densities of the above-mentioned protein measured via ImageJ and related to the housekeepers lamin B2 (68 kDa) and GAPDH (37 kDa). ( c ) Representative pictures of cortical and medullary kidney sections from control and aldosterone-infused animals stained with an antibody against Nrf2. Examples of positive stained areas are marked with black arrows. Percentage of Nrf2-positive stained areas in cortex ( d ) and medulla ( e ). For the quantification of positive Nrf2 areas, 10 visual fields of cortical and 3–5 visual fields of medullary kidney sections were analyzed per animal via ImageJ. Aldo: aldosterone, GAPDH: glyceraldehyde 3-phosphate dehydrogenase, Nrf2: nuclear factor erythroid 2-related factor 2, Sulf. sulforaphane, WT: wild-type. Data are shown as mean + SEM, n = 7–8. * p ≤ 0.05 vs. WT-C, # p ≤ 0.05 vs. WT-Sulf, ° p ≤ 0.05 vs. WT-Aldo, ^ p ≤ 0.05 vs. Nrf2ꜛ-C.

Article Snippet: Membranes were incubated with specific primary antibodies against Bach1 (abx322188, Abbexa, Cambridge, UK), FYN (ab184276, abcam, Cambridge, UK), γGCLC (PA1492, BosterBio, Pleasanton, CA, USA), GSK3β (#9315, Cell Signaling, Herts, UK), pGSK3β (#9336, Cell Signaling, Herts, UK), HO1 (ab13243, abcam, Cambridge, UK), Keap1 (ab227828, abcam, Cambridge, UK), MafK (GTX129240, GeneTex, Irvine, CA, USA), NQO1 (ab34173, abcam, Cambridge, UK), Nrf2 (sc-722, Santa Cruz Biotechnology, Dallas, TX, USA), pNrf2 (ab76026, abcam, Cambridge, UK), SOD1 (GTX100554, GeneTex, Irvine, CA, USA), TrxR1 (GTX108727, GeneTex, Irvine, CA, USA) and, as housekeepers, α-tubulin (sc-5286, Santa Cruz Biotechnology, Dallas, TX, USA), GAPDH (#2118, Cell Signaling, Herts, UK) and lamin B2 (#2328, Cell Signaling, Herts, UK) overnight at 4 °C, followed by an incubation with HRP-conjugated secondary antibodies for 2.5 h at room temperature.

Techniques: Control, Western Blot, Expressing, Staining

Journal: Antioxidants

Article Title: Nrf2 Activation Does Not Protect from Aldosterone-Induced Kidney Damage in Mice

doi: 10.3390/antiox12030777

Figure Lengend Snippet: Localization of the activated transcription factor Nrf2. Representative pictures of the Western blots of the expression of total Nrf2 phosphorylated at Ser40 (pNrf2, 110 kDa) in ( a ) nuclear and ( b ) cytosolic extracts of kidneys of mice as well as the quantification of band densities of the above-mentioned protein measured via ImageJ and related to the housekeepers lamin B2 (68 kDa) and GAPDH (37 kDa). Percentage of pNrf2-positive stained nuclei in cortex ( c ) and medulla ( d ). For the quantification of positive Nrf2 nuclei, 10 visual fields of cortical and 3–5 visual fields of medullary kidney sections were analyzed per animal via ImageJ. ( e ) Representative images of double stained kidney sections used for the localization of pNrf2 in kidney cells. Double staining on paraffin-embedded kidney sections was carried out using antibodies against pNrf2 (brown staining) and against calbindin (purple staining), a marker for distal tubule and early collecting duct cells. Examples of pNrf2-positive stained nuclei are indicated by black arrows; white arrows indicate the corresponding section of the tubulus system. Proximal tubules were identified by the presence of the brush border, whereas glomeruli were identified by their capillary tuft (blue circles). Distal tubules and the early collecting duct were visualized by positive calbindin staining. The late collecting duct was identified by the absence of positive calbindin staining and brush border. ( f – i ) Quantification of pNrf2-positive nuclei in the four kidney structures related to the number of nuclei in regions positive for the specific kidney cell identifiers in 10 visual fields. For the quantification in the glomerulus, 50 glomeruli were evaluated. Aldo: aldosterone, C: control, Nrf2: nuclear factor erythroid 2-related factor 2, Sulf: sulforaphane, WT: wild type. Data are shown as mean + SEM, n = 7–8. * p ≤ 0.05 vs. WT-C, # p ≤ 0.05 vs. WT-Sulf, ° p ≤ 0.05 vs. WT-Aldo, ^ p ≤ 0.05 vs. Nrf2ꜛ-C.

Article Snippet: Membranes were incubated with specific primary antibodies against Bach1 (abx322188, Abbexa, Cambridge, UK), FYN (ab184276, abcam, Cambridge, UK), γGCLC (PA1492, BosterBio, Pleasanton, CA, USA), GSK3β (#9315, Cell Signaling, Herts, UK), pGSK3β (#9336, Cell Signaling, Herts, UK), HO1 (ab13243, abcam, Cambridge, UK), Keap1 (ab227828, abcam, Cambridge, UK), MafK (GTX129240, GeneTex, Irvine, CA, USA), NQO1 (ab34173, abcam, Cambridge, UK), Nrf2 (sc-722, Santa Cruz Biotechnology, Dallas, TX, USA), pNrf2 (ab76026, abcam, Cambridge, UK), SOD1 (GTX100554, GeneTex, Irvine, CA, USA), TrxR1 (GTX108727, GeneTex, Irvine, CA, USA) and, as housekeepers, α-tubulin (sc-5286, Santa Cruz Biotechnology, Dallas, TX, USA), GAPDH (#2118, Cell Signaling, Herts, UK) and lamin B2 (#2328, Cell Signaling, Herts, UK) overnight at 4 °C, followed by an incubation with HRP-conjugated secondary antibodies for 2.5 h at room temperature.

Techniques: Western Blot, Expressing, Staining, Double Staining, Marker, Control

Journal: Antioxidants

Article Title: Nrf2 Activation Does Not Protect from Aldosterone-Induced Kidney Damage in Mice

doi: 10.3390/antiox12030777

Figure Lengend Snippet: Expression of proteins important for the function of active Nrf2 and of proteins regulating Nrf2 activity in Aldo-infused WT and Nrf2ꜛ mice and control mice after 28 days of treatment. Representative pictures of Western blots and the quantification of band densities measured via ImageJ and related to the nuclear protein lamin B2 (68 kDa) or, in the case of pGSK3β, to non-phosphorylated GSK3β (46 kDa) of ( a ) nuclear localized MafK (18 kDa), ( b ) nuclear localized Bach1 (81 kDa), ( c ) GSK3β (46 kDa) localized in the cytosol, ( d ) nuclear localized GSK3β (46 kDa), ( e ) total phosphorylated GSK3β (46 kDa) and ( f ) nuclear localized FYN (60 kDa) are shown. Aldo: aldosterone, Bach1: BTB and CNC homology 1, basic leucine zipper transcription factor 1, C: control, FYN: Src family tyrosine kinase, GSK3β: glyceraldehyde 3-phosphate dehydrogenase, MafK: musculoaponeurotic fibrosarcoma K, Sulf: sulforaphane, WT: wild-type. Data are shown as mean + SEM, n = 7–8. * p ≤ 0.05 vs. WT-C, # p ≤ 0.05 vs. WT-Sulf, ° p ≤ 0.05 vs. WT-Aldo.

Article Snippet: Membranes were incubated with specific primary antibodies against Bach1 (abx322188, Abbexa, Cambridge, UK), FYN (ab184276, abcam, Cambridge, UK), γGCLC (PA1492, BosterBio, Pleasanton, CA, USA), GSK3β (#9315, Cell Signaling, Herts, UK), pGSK3β (#9336, Cell Signaling, Herts, UK), HO1 (ab13243, abcam, Cambridge, UK), Keap1 (ab227828, abcam, Cambridge, UK), MafK (GTX129240, GeneTex, Irvine, CA, USA), NQO1 (ab34173, abcam, Cambridge, UK), Nrf2 (sc-722, Santa Cruz Biotechnology, Dallas, TX, USA), pNrf2 (ab76026, abcam, Cambridge, UK), SOD1 (GTX100554, GeneTex, Irvine, CA, USA), TrxR1 (GTX108727, GeneTex, Irvine, CA, USA) and, as housekeepers, α-tubulin (sc-5286, Santa Cruz Biotechnology, Dallas, TX, USA), GAPDH (#2118, Cell Signaling, Herts, UK) and lamin B2 (#2328, Cell Signaling, Herts, UK) overnight at 4 °C, followed by an incubation with HRP-conjugated secondary antibodies for 2.5 h at room temperature.

Techniques: Expressing, Activity Assay, Control, Western Blot