student’s two-tailed t-test analysis  (GraphPad Software Inc)

Journal: Nature Biomedical Engineering

Article Title: Nitroglycerin-responsive gene switch for the on-demand production of therapeutic proteins

doi: 10.1038/s41551-025-01350-7

Figure Lengend Snippet: a , Schematic illustration of the in vivo experiment conducted to examine the performance of engineered cells in mice. b , Alginate–PLL–alginate encapsulated HEK hNORM1 cells (5 × 10 6 cells in total) were first implanted subcutaneously into C57BL/6 mice . At 24 h after device implantation, different dosages of NG patches were topically applied above the cell implant. NLuc levels in the blood were measured 24 h following the drug administration. Data are presented as mean ± s.e.m. ( n = 5), and P values were calculated using a two-tailed, unpaired Student’s t -test. c , Immunoblotting for ALDH2 of parental HEK-293 WT, HEK hNORM1 and HEK hNORM2 . α-Actinin was used as a loading control. d , NLuc levels in blood of mice as described in b using HEK hNORM2 designer cells instead of HEK hNORM1 , and an NG-patch dose of 130 µg per 24 h. Data are presented as mean ± s.e.m. ( n = 5), and P values were calculated using a two-tailed, unpaired Student’s t -test. e , NO levels in skin tissue of mice following the administration of an NG patch. Encapsulated HEK-293 WT, HEK hNORM1 or HEK hNORM2 cells were subcutaneousinfluencing deep tissues.ly delivered into C57BL/6 mice, and an NG patch (130 µg per 24 h) was applied on top of the implantation site as described in b . NO levels in skin tissue beneath the NG patches were assessed at 8 h intervals. Data are presented as mean ± s.e.m. ( n = 4). Statistical significance was analysed by two-way ANOVA and P values were calculated using Tukey’s multi-comparison tests. f , HEK hNORM2 -containing microcapsules were implanted either subcutaneously or intraperitoneally into four groups of C57BL/6 mice (5 × 10 6 cells per mouse). At 2 h following the implantation, NG patches (130 µg per 24 h) were applied to the back and NLuc in the blood was quantified after 24 h. WT mice implanted with parental HEK-293 cells were used as a negative control. Data are presented as mean ± s.e.m. ( n = 5), and P values were calculated using a two-tailed, unpaired Student’s t -test. IP, intraperitoneal; SQ, subcutaneous; NS, not significant. g , In vivo evaluation of reversibility using NG patches. HEK hNORM2 -containing microcapsules were subcutaneously implanted into mice at day 0 using 5 × 10 6 cells per mouse. NG patches (130 µg per 24 h) were applied above the implants on days 1, 3 and 5 (for 24 h each time), while on days 2, 4, 6, 7, 8 and 9, mice were kept without an NG patch. Blood samples were taken every day at the same time just before patch application and removal. Data are presented as mean ± s.e.m. ( n = 4). All data shown are biological replicates. Source data are provided as a Source Data file.

Article Snippet: The statistical significance of differences among groups was evaluated with a two-tailed, unpaired Student’s t -test or analysis of variance (ANOVA) using GraphPad Prism.

Techniques: In Vivo, Two Tailed Test, Western Blot, Control, Comparison, Negative Control

Journal: Nature Biomedical Engineering

Article Title: Nitroglycerin-responsive gene switch for the on-demand production of therapeutic proteins

doi: 10.1038/s41551-025-01350-7

Figure Lengend Snippet: a , ELISA quantification of active GLP-1 levels in the supernatants of HEK hNORM3 cells following DETA NONOate (100 µM) treatment for 24 h. Data are presented as mean ± s.d. ( n = 6), and P values were calculated using a two-tailed, unpaired Student’s t -test. b , c , Long-term performance and anti-glycaemic effect of hNORM in db/db diabetic mice. HEK hNORM3 cell-containing microcapsules (5 × 10 6 cells per mouse) were subcutaneously implanted into db/db mice at days 0 and 14, and NG patches were topically applied in the vicinity of the implant. NG patches (130 µg per 24 h) were applied once every 2 days starting from day 0 until the end of the experiment. Plasma GLP-1 ( b ) and fasting glucose ( c ) levels were analysed every 3 days for 35 days. Data are presented as mean ± s.e.m. ( n = 4), and P values were calculated using a two-tailed, unpaired Student’s t -test. d , e , MAP ( d ) and heart rate ( e ) of mice described in the previous experiment measured at 7 day intervals. f , g , MAP ( f ) and heart rate ( g ) of WT mice harbouring subcutaneous cell implants that contain either HEK-293 WT or HEK hNORM3 cells. NG patches (130 µg per 24 h) were applied at 24 h after the implantation, and cardiovascular parameters were measured at times 0 h and 24 h following the patch application. Data are presented as mean ± s.e.m. ( n = 4). Statistical significance was analysed by two-way ANOVA, and P values were calculated using Tukey’s multi-comparison tests. All data shown are biological replicates. Source data are provided as a Source Data file.

Article Snippet: The statistical significance of differences among groups was evaluated with a two-tailed, unpaired Student’s t -test or analysis of variance (ANOVA) using GraphPad Prism.

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, Clinical Proteomics, Comparison