Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet: Adaptable expression ratios of the creatine transporters CRT1 and MCT12 in HEK293 SLC6A8 −/− cells (A) Scheme of the CRT1 3xHA -P2A-K n -MCT12 2xmyc construct. (B) Immunoblot analysis of CRT1 3xHA -P2A-K n -MCT12 2xmyc . (C) Ratio of MCT12 2xmyc and CRT1 3xHA expression as determined by immunoblotting ( n = 5). Data are represented as mean ± SEM. (D) Immunoblot analysis of CRT1 3xHA -P2A-K n -MCT12 2xmyc . (E) Ratio of MCT12 2xmyc and CRT1 3xHA expression as determined by immunoblotting ( n = 4). Data are represented as mean ± SEM. See also .

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Expressing, Construct, Western Blot, Analysis

Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet: Relative cell surface expression of the creatine transporters CRT1 and MCT12 in HEK293 SLC6A8 −/− cells (A) Scheme of the CRT1 eGFP -P2A-K 0 -MCT12 mCherry (a) and MCT12 mCherry -P2A-K 0 -CRT1 eGFP construct (b). (B) Ratio of mCherry and eGFP expression in eGFP-P2A-K 0 -mCherry (K 0 , as a control), CRT1 eGFP -P2A-K 0 -MCT12 mCherry , and MCT12 mCherry -P2A-K 0 -CRT1 eGFP constructs as determined by flow cytometry ( n = 6). Data are represented as mean ± SEM. (C) Live-cell confocal microscopy of HEK293 SLC6A8 −/− cells expressing CRT1 eGFP -P2A-K 0 -MCT12 mCherry and MCT12 mCherry -P2A-K 0 -CRT1 eGFP . The plasma membrane was stained with CellMask Deep Red. Scale bars, 20 μm. (D) Analysis of cell surface expression of CRT1 eGFP and MCT12 mCherry . n = 30 (CRT1 eGFP -P2A-K 0 -MCT12 mCherry ); n = 28 (MCT12 mCherry -P2A-K 0 -CRT1 eGFP ).

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Expressing, Construct, Control, Flow Cytometry, Confocal, Microscopy, Clinical Proteomics, Membrane, Staining, Analysis

Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet:

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Cell-Signaling, Virus, Recombinant, Clinical Proteomics, Membrane, Staining, Cloning, Software

Journal: iScience

Article Title: Sulfonylurea receptor 1 deficiency alleviates acute lung injury by activating PARP1 in epithelial cells

doi: 10.1016/j.isci.2026.117627

Figure Lengend Snippet: SUR1 protein levels were elevated in the bronchoalveolar lavage fluid (BALF), but not the serum, of sepsis-related ALI patients and CLP-induced ALI mouse models (A) Dot blot analysis of BALF and serum collected from sepsis-related ALI patients or controls. NC (negative control), PBS; PC (positive control), whole-cell protein lysates of HEK293T cells. Control, patients who underwent abdominal surgery without ALI or bacterial infection; ALI, sepsis-associated ALI. Red circle, site of PBS drops. (B) Cytokine levels in BALF and serum of patients determined by a cytokine screening assay. n = 3 for control; n = 9 for ALI. (C) Dot blot analysis of BALF and serum from CLP-induced ALI mice ( n = 6). (D–F) Western blot and RT-qPCR analyses of lung tissue samples from CLP-induced ALI mice. Data are presented as mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, no significance. Data in (A, B, C, and F) were analyzed using Welch’s t test, and data in (E) were analyzed using an unpaired two-tailed Student’s t test. Representative images are shown.

Article Snippet: Human embryonic kidney cell line HEK293T , American Type Culture Collection , N/A.

Techniques: Dot Blot, Analysis, Negative Control, Positive Control, Control, Infection, Screening, Western Blot, Quantitative RT-PCR, Tissue, Two Tailed Test

Journal: iScience

Article Title: Sulfonylurea receptor 1 deficiency alleviates acute lung injury by activating PARP1 in epithelial cells

doi: 10.1016/j.isci.2026.117627

Figure Lengend Snippet: SUR1 interacted with PARP1 to suppress DNA damage repair and induce apoptosis of bronchial epithelial cells (A) Co-immunoprecipitation and western blot analyses of HEK293T cells transfected with FLAG-SUR1 plasmids (left) or human bronchial epithlial (HBE) cells (right). (B) Immunofluorescence staining of HBE cells. Scale bars, 25 μm. (C) Homologous recombination reporter assay in U2OS-DRGFP cells transfected with SUR1 siRNA, followed by flow cytometry analysis. (D) Western blot analysis of HBE cells transfected with SUR1 siRNAs with or without 20 ng/mL TNF-α or 50 μg/mL LPS treatment for 1 h. (E) ROS levels in HBE cells with SUR1 knockdown, treated with or without TNF-α 20 ng/mL for 3 h. (F) Apoptosis assay by flow cytometry in HBE cells transfected with SUR1 siRNAs and treated with TNF-α. Data are presented as mean ± SD ( n = 3). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Co-localization in (B) was assessed using Pearson’s correlation coefficient. Data in (C–F) were analyzed using two-way ANOVA followed by Tukey’s multiple-comparisons test. Representative images are shown.

Article Snippet: Human embryonic kidney cell line HEK293T , American Type Culture Collection , N/A.

Techniques: Co-Immunoprecipitation Assay, Western Blot, Transfection, Immunofluorescence, Staining, Homologous Recombination, Reporter Assay, Flow Cytometry, Analysis, Knockdown, Apoptosis Assay

Journal: iScience

Article Title: Sulfonylurea receptor 1 deficiency alleviates acute lung injury by activating PARP1 in epithelial cells

doi: 10.1016/j.isci.2026.117627

Figure Lengend Snippet:

Article Snippet: Human embryonic kidney cell line HEK293T , American Type Culture Collection , N/A.

Techniques: Cell-Signaling, Recombinant, Transfection, Caspase-3 Activity Assay, Protein Extraction, Knock-Out, Sequencing, Control, Plasmid Preparation, Software, Analysis