Journal: bioRxiv

Article Title: Wnts are endothelial cell-derived PKD1/PKD2-dependent autocrine/paracrine vasodilators

doi: 10.64898/2026.03.17.712518

Figure Lengend Snippet: (A) Representative RT-PCR showing that mRNA for PKD1 is present in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl but absent in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl :Cdh5(PAC)-CreERT2 mice. Representative of 5 independent experiments for each genotype. (B) Representative Western blots illustrating PKD1, PKD2, AT1, eNOS, Fzd-7 and actin proteins in mesenteric arteries of Pkd1 fl/fl and Pkd1 ecKO mice. (C) Mean data from experiments shown in panel B. Significance was assessed using Student t-tests, n=5 independent mesenteric arterial lysates for each genotype and each protein. (D) Immunofluorescence images (representative of three mesenteric arteries from three mice for each genotype) illustrating that PKD1 (Alexa Fluor 546) is abolished in endothelial cells of en face mesenteric arteries from Pkd1 ecKO mice. CD31 (Alexa Fluor 488) and DAPI are also shown. Scale bars, 50 μm. (E) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt9b (3 μg/ml) applied in continuous flow in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (F) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt9b (3 µg/ml) applied under continuous flow (10 dyn/cm 2 ) in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (G) Mean data from experiments shown in panels E and F, and the modulation of flow and Wnt9b-mediated vasodilation by SRI37892 (Fzd-7 Inh, 5 µM). Significance was assessed using a two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=10 arteries from 7 mice of each genotype for flow, flow+ Wnt9b and Wnt9b. n=7 arteries from 5 mice for Fzd-7 Inh+flow and Fzd-7 Inh+flow+Wnt9b. n=5 arteries from 5 mice for each genotype for flow + boiled Wnt9b. (H) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt5a (3 µg/ml). (I) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt5a (3 µg/ml). (J) Mean data from experiments shown in panels H and I and the modulation of flow and Wnt5a-induced vasodilation by SRI37892 (Fzd-7 Inh, 5 μM). Significance was assessed using two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=8 arteries from 6 mice from Pkd1 fl/fl for flow, flow+ Wnt5a. n=5 arteries from 5 mice from Pkd1 ecKO for flow, flow+ Wnt5a. n=5 arteries from 5 mice of each genotype for flow + boiled Wnt5a. n=8 arteries from 6 mice from Pkd1 fl/fl for flow+ Fzd-7 Inh and flow+ Wnt5a+ Fzd-7 Inh.

Article Snippet: Membranes were blocked with 5% milk or 5% BSA and incubated with one of the following primary antibodies: PKD1 (Polycystic Kidney Disease Research Resource Consortium, Baltimore), PKD2 (Alomone), eNOS (Abcam), p-eNOS (Cell Signaling), Wnt9b (R&D Systems), Wnt5a (R&D Systems), AT1 receptor (Alomone) or actin (Cell Signaling) overnight at 4°C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Western Blot, Immunofluorescence

Journal: Molecular Medicine Reports

Article Title: ULK1 activates NCOA4-mediated ferritinophagy via the Beclin1/VPS34 complex in cardiomyocyte hypertrophy

doi: 10.3892/mmr.2026.13826

Figure Lengend Snippet: ULK1 expression is elevated in hypertrophic cardiac tissues and cardiomyocytes. (A) C57BL/6J mice at 8-weeks-old were subjected to sham or TAC surgery. Representative whole heart images (scale bar, 0.5 cm), M-mode echocardiography, H&E staining (scale bar, 100 µm) and WGA staining (scale bar, 50 µm) of heart tissue. Assessment of (B) HW/BW, (C) LVW/BW and (D) HW/TL (n=6). Statistical graphs for (E) LVEF (%), (F) FS (%), (G) IVS, d (mm) and (H) LVPW, d (mm) (n=6). (I) Cardiomyocyte cross-sectional area acquired from WGA staining. Cells were measured from different microscopic fields of 6 samples in each group. The mRNA levels of (J) BNP and (K) β-MHC in the hearts from sham or TAC-surgery mice (n=6). The mRNA levels of (L) BNP and (M) β-MHC in HL-1 cells treated with Ang II (n=3). (N) Stained cells and (O) quantification of cell surface area of HL-1 cells treated with Ang II. Scale bar, 50 µm (n=3). Western blot analysis and summarized data demonstrating (P) Beclin1 and (Q) VPS34 protein expression in HL-1 cells treated with Ang II (n=3). (R) Western blot analysis and summarized data demonstrating ULK1 protein levels in the hearts obtained after sham or TAC surgery (n=6). (S) Western blot analysis and summarized data demonstrating ULK1 protein levels in the cardiomyocytes stimulated with Ang II (n=6). All data are presented as mean ± SEM. *P < 0.05 and **P<0.01 vs. sham or control group. ULK1, serine/threonine protein kinase ULK1; TAC, transverse aortic constriction; HW, heart weight; BW, body weight; LVW, left ventricular weight; TL, tibia length; LVEF, left ventricular ejection fraction; FS, fractional shortening; d, diastole; IVS, interventricular septal thickness; LVPW, left ventricular posterior wall thickness; CSA, cross-sectional area; BNP , brain natriuretic peptide; β-MHC , β-myosin heavy chain; α-SMA, α smooth muscle actin; Ang II, angiotensin II; VPS34, PI3K catalytic subunit type 3; WGA, wheat germ agglutinin.

Article Snippet: The Ang II AT1-receptor candesartan (CS; cat. no. HY-B0205) was purchased from MedChemExpress.

Techniques: Expressing, Staining, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: ULK1 activates NCOA4-mediated ferritinophagy via the Beclin1/VPS34 complex in cardiomyocyte hypertrophy

doi: 10.3892/mmr.2026.13826

Figure Lengend Snippet: Mechanisms of ULK1 activating NCOA4-mediated ferritinophagy via Beclin1/VPS34 complex in cardiomyocyte hypertrophy. This figure was created using Figdraw ( www.figdraw.com ). ULK1, serine/threonine protein kinase ULK1; NCOA4, nuclear receptor coactivator 4; VPS34, PI3K catalytic subunit type 3; LC3, microtubule-associated protein 1 light chain 3; TAC, transverse aortic constriction; Ang II, angiotensin II; ROS, reactive oxygen species.

Article Snippet: The Ang II AT1-receptor candesartan (CS; cat. no. HY-B0205) was purchased from MedChemExpress.

Techniques:

Journal: Biomedicines

Article Title: Long-Term Alterations of Renal Microvasculature in Rats Following Maternal PM2.5 Exposure: Vitamin D Effects

doi: 10.3390/biomedicines13051166

Figure Lengend Snippet: Figure 5. Intrarenal RAS signaling in adult offspring kidneys: (a–d) renin. (e–h) ACE. (i) AT1. Western blot analysis showed increased renin (a) and ACE (e) activities in the kidneys of offspring rats exposed to maternal PM2.5 compared to controls. Maternal vitamin D supplementation significantly reduced intrarenal ACE activity (e). Immunohistochemically, renin expression was observed in the interstitium and juxtaglomerular cells of offspring kidneys from the PM ((c), arrows) and PV groups ((d), arrows), whereas it was rarely detected in control offspring kidneys (b). ACE was also expressed extensively throughout the brush border of tubular cells in kidneys of offspring exposed to maternal PM2.5 ((g), arrows). Maternal vitamin D supplementation markedly reduced intrarenal ACE expression in the adult offspring rats (h). No differences were observed in intrarenal AT1 activity among the groups (i) (* p < 0.05, Cont vs. PM; ** p < 0.05, Cont vs. PV; † p < 0.05, PM vs. PV) (200×, bar = 50 µm) (n = 3–6/group). Cont, control group; PM, PM group; PV, PV group; ns, not significant.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies against vitamin D receptor (VDR) (Abcam, Cambridge, MA, USA), Klotho (Invitrogen, Carlsbad, CA, USA), cytochrome P450 mixed-function oxidase (CYP)27B1 (Abcam), CYP24A1 (Invitrogen), renin (Santa Cruz biotechnology, Santa Cruz, CA, USA), angiotensin-converting enzyme (ACE) (Invitrogen), angiotensin II receptor type 1 (AT1) (Invitrogen), VEGF-A (Santa Cruz Biotechnology), VEGFR2 (Santa Cruz Biotechnology), hypoxia-inducible factor-1 alpha (HIF-1α) (Santa Cruz Biotechnology), Ang-1 (Santa Cruz Biotechnology), Tie-2 (Santa Cruz Biotechnology), Ang-2 (Santa Cruz Biotechnology), and thrombospondin (TSP)-1 (Santa Cruz Biotechnology).

Techniques: Western Blot, Activity Assay, Expressing, Control