unc5cl  (Cell Signaling Technology Inc)

Journal: Cell Reports Medicine

Article Title: Phosphoglycerate kinase 1 contributes to diabetic kidney disease through enzyme-dependent and independent manners

doi: 10.1016/j.xcrm.2025.102241

Figure Lengend Snippet: PGK1 activated UNC5CL-mediated inflammatory response in DKD (A) Heatmap showing the differentially expressed proteins in HK-2 cells ( n = 3). (B) Volcano plot showing the differentially expressed proteins in HK-2 cells. (C and D) Representative blots and quantitative analysis of GPX1 ( n = 4). (E) Effects of silencing UNC5CL on the protein expression of collagen I, 12-Lox, and IL-1β ( n = 3). (F) Effects of silencing PGK1 on the protein expression of UNC5CL, IκBα, P-IκBα, P-NF-κB, and P-JNK ( n = 3). (G) Effects of silencing PGK1 on the mRNA levels of CXCL1 , CXCL2 , CXCL3 , IL8 , CCL20 , TNFAIP3 , PLA2G4C , and TNFRSF9 ( n = 4). Data are represented as mean ± SD. Statistical significance was determined by unpaired Student’s t test or ANOVA. ∗ p < 0.05 vs. NG, control (Con) or HG + Con small interfering RNA (siRNA), † p < 0.05 vs. Control (Con) siRNA. See also .

Article Snippet: Anti-UNC5CL antibody , Proteintech , Cat# 13322-1-AP, RRID: AB_2877938.

Techniques: Expressing, Control, Small Interfering RNA

Journal: Cell Reports Medicine

Article Title: Phosphoglycerate kinase 1 contributes to diabetic kidney disease through enzyme-dependent and independent manners

doi: 10.1016/j.xcrm.2025.102241

Figure Lengend Snippet: Aldh1l1 bridges the link between PGK1 and UNC5CL in the development of DKD (A) Aldh1l1 was identified to interact with PGK1. (B) Representative blots and quantitative analysis of Aldh1l1 ( n = 4). (C–E) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in NG- or HG-incubated HK-2 cells ( n = 4). (F–H) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in kidneys from control and T1D-DKD mice ( n = 4). (I–K) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in kidneys from control and T2D-DKD mice ( n = 4). (L) Effects of Aldh1l1 small interfering RNA (siRNA) on the complex formation of PGK1/UNC5CL ( n = 4). (M and N) Effects of Aldh1l1 siRNA on the impact of PGK1 overexpression (OE) on the UNC5CL-mediated inflammation ( n = 4). Data are represented as mean ± SD. Statistical significance was determined by unpaired Student’s t test.∗ p < 0.05 vs. NG or control (Con). See also and .

Article Snippet: Anti-UNC5CL antibody , Proteintech , Cat# 13322-1-AP, RRID: AB_2877938.

Techniques: Incubation, Control, Small Interfering RNA, Over Expression

Journal: Cell Reports Medicine

Article Title: Phosphoglycerate kinase 1 contributes to diabetic kidney disease through enzyme-dependent and independent manners

doi: 10.1016/j.xcrm.2025.102241

Figure Lengend Snippet: PGK1 activated UNC5CL-mediated inflammatory response in DKD (A) Heatmap showing the differentially expressed proteins in HK-2 cells ( n = 3). (B) Volcano plot showing the differentially expressed proteins in HK-2 cells. (C and D) Representative blots and quantitative analysis of GPX1 ( n = 4). (E) Effects of silencing UNC5CL on the protein expression of collagen I, 12-Lox, and IL-1β ( n = 3). (F) Effects of silencing PGK1 on the protein expression of UNC5CL, IκBα, P-IκBα, P-NF-κB, and P-JNK ( n = 3). (G) Effects of silencing PGK1 on the mRNA levels of CXCL1 , CXCL2 , CXCL3 , IL8 , CCL20 , TNFAIP3 , PLA2G4C , and TNFRSF9 ( n = 4). Data are represented as mean ± SD. Statistical significance was determined by unpaired Student’s t test or ANOVA. ∗ p < 0.05 vs. NG, control (Con) or HG + Con small interfering RNA (siRNA), † p < 0.05 vs. Control (Con) siRNA. See also .

Article Snippet: After blocking with 5% non-fat dry milk (5g skim milk powder in 100mL TBST) at 37°C for 1 h, the membranes were incubated overnight at 4°C with the following primary antibodies: PGK1 (17811-1-AP, Proteintech, Rosemont, USA), 12-Lox (NBP2-46512, Novus, Centennial, CO, USA), P-p47 phox (Ser328, ab111855, Abcam, Waltham, MA, USA), Collagen I (ab6308, Abcam, Waltham, MA, USA), cleaved-caspase 3 (9664, Cell Signaling Technology, Danvers, MA, USA), GPX1 (29329-1-AP, Proteintech, Rosemont, USA), β-actin (81115-1-RR, Proteintech, Rosemont, USA), NLRP3 (15101, Cell Signaling Technology, Danvers, MA, USA), ASC (10500-1-AP, Proteintech, Rosemont, USA), Caspase-1 (ab74279, Abcam, Waltham, MA, USA), UNC5CL (13322-1-AP, Proteintech, Rosemont, USA), IκBα (9242, Cell Signaling Technology, Danvers, MA, USA), P-IκBα (Ser32, 2859, Cell Signaling Technology, Danvers, MA, USA), p-NF-κB (Ser536, 3033, Cell Signaling Technology, Danvers, MA, USA), NF-κB (8242, Cell Signaling Technology, Danvers, MA, USA), p-JNK (Thr183/Tyr185, 4668, Cell Signaling Technology, Danvers, MA, USA), JNK (9252, Cell Signaling Technology, Danvers, MA, USA), Aldh1l1 (17390-1-AP, Proteintech, Rosemont, USA), IRAK1 (10478-2-AP, Proteintech, Rosemont, USA), IRAK4 (4363, Cell Signaling Technology, Danvers, MA, USA), TRAF6 (14424-1-AP, Proteintech, Rosemont, USA), PAX5 (8970, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Control, Small Interfering RNA

Journal: Cell Reports Medicine

Article Title: Phosphoglycerate kinase 1 contributes to diabetic kidney disease through enzyme-dependent and independent manners

doi: 10.1016/j.xcrm.2025.102241

Figure Lengend Snippet: Aldh1l1 bridges the link between PGK1 and UNC5CL in the development of DKD (A) Aldh1l1 was identified to interact with PGK1. (B) Representative blots and quantitative analysis of Aldh1l1 ( n = 4). (C–E) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in NG- or HG-incubated HK-2 cells ( n = 4). (F–H) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in kidneys from control and T1D-DKD mice ( n = 4). (I–K) CoIP results showing the complex formation of PGK1/UNC5CL/Aldh1l1 in kidneys from control and T2D-DKD mice ( n = 4). (L) Effects of Aldh1l1 small interfering RNA (siRNA) on the complex formation of PGK1/UNC5CL ( n = 4). (M and N) Effects of Aldh1l1 siRNA on the impact of PGK1 overexpression (OE) on the UNC5CL-mediated inflammation ( n = 4). Data are represented as mean ± SD. Statistical significance was determined by unpaired Student’s t test.∗ p < 0.05 vs. NG or control (Con). See also and .

Article Snippet: After blocking with 5% non-fat dry milk (5g skim milk powder in 100mL TBST) at 37°C for 1 h, the membranes were incubated overnight at 4°C with the following primary antibodies: PGK1 (17811-1-AP, Proteintech, Rosemont, USA), 12-Lox (NBP2-46512, Novus, Centennial, CO, USA), P-p47 phox (Ser328, ab111855, Abcam, Waltham, MA, USA), Collagen I (ab6308, Abcam, Waltham, MA, USA), cleaved-caspase 3 (9664, Cell Signaling Technology, Danvers, MA, USA), GPX1 (29329-1-AP, Proteintech, Rosemont, USA), β-actin (81115-1-RR, Proteintech, Rosemont, USA), NLRP3 (15101, Cell Signaling Technology, Danvers, MA, USA), ASC (10500-1-AP, Proteintech, Rosemont, USA), Caspase-1 (ab74279, Abcam, Waltham, MA, USA), UNC5CL (13322-1-AP, Proteintech, Rosemont, USA), IκBα (9242, Cell Signaling Technology, Danvers, MA, USA), P-IκBα (Ser32, 2859, Cell Signaling Technology, Danvers, MA, USA), p-NF-κB (Ser536, 3033, Cell Signaling Technology, Danvers, MA, USA), NF-κB (8242, Cell Signaling Technology, Danvers, MA, USA), p-JNK (Thr183/Tyr185, 4668, Cell Signaling Technology, Danvers, MA, USA), JNK (9252, Cell Signaling Technology, Danvers, MA, USA), Aldh1l1 (17390-1-AP, Proteintech, Rosemont, USA), IRAK1 (10478-2-AP, Proteintech, Rosemont, USA), IRAK4 (4363, Cell Signaling Technology, Danvers, MA, USA), TRAF6 (14424-1-AP, Proteintech, Rosemont, USA), PAX5 (8970, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Incubation, Control, Small Interfering RNA, Over Expression

Journal: Cell Death and Differentiation

Article Title: The death domain-containing protein Unc5CL is a novel MyD88-independent activator of the pro-inflammatory IRAK signaling cascade

doi: 10.1038/cdd.2011.147

Figure Lengend Snippet: Tissue distribution and subcellular localization of Unc5CL. (A) Unc5CL-specific western blot of whole tissue lysates from the indicated tissues or overexpressed untagged Unc5CL as marker. (B) Relative Unc5CL mRNA expression in epithelial cell preparations from consecutive small intestinal (1–6) or colonic segments. Data represent the mean values ±S.D. of technical triplicates. (C) Unc5CL-specific western blot of intestinal (1–6) and colonic epithelial samples shown in (B). (D) Unc5CL-specific western blot of lysates from CaCo-2 cells stably expressing mock (scr sh) or Unc5CL-specific shRNAs (sh1–sh5). (E) Proteins from CaCo-2 cells were subjected to TX-114 phase separation. Fractions were analyzed by western blot using the indicated antibodies. CytoC, 14 kDa Cytochrome C, I, detergent-insoluble proteins, D, detergent phase, amphiphilic integral membrane proteins, A, aqueous phase, hydrophilic proteins. (F) z-stack reconstruction following confocal microscopy of overexpressed C-terminally FLAG-tagged Unc5CL (Unc5CL) or Unc5CL lacking the transmembrane domain (ΔTM) in CaCo-2 cells using FLAG-specific antibodies. (G) (a–c) Immunofluorescent staining of Unc5CL on cryosections from OCT embedded uterus using Unc5CL-specific antibodies. Blue: nuclear DAPI staining. (a, b) Green: Unc5CL, (c) isotype control (FLAG). (A, D and E) Asterisks mark unspecific bands

Article Snippet: Lentiviral pLKO.1 plasmids expressing Unc5CL-specific shRNAs were obtained from Sigma. siRNAs were obtained from Ambion (Rotkreuz, Switzerland).

Techniques: Western Blot, Marker, Expressing, Stable Transfection, Confocal Microscopy, Staining