Journal: Scientific Reports

Article Title: Dexamethasone enhances intestinal glucose absorption and TMPRSS2 expression with implications for hyperglycaemia and infection risk

doi: 10.1038/s41598-025-22312-8

Figure Lengend Snippet: Inflammation upregulates SGLT1, ACE2 and TMPRSS2 in Caco-2/TC7 cells. ( A ) Graphical illustration of the experimental protocol for the standard and inflamed Caco-2/TC7 cell models. Cells were differentiated in 25 mM glucose until day 7, then in 5.5 mM glucose until day 14. Cells were differentiated for 14 days from confluence and treated with dexamethasone (DEX) or 0.1% (v/v) DMSO (vehicle control) up to twice daily for 60 h, 24 h, or 4 h. For the inflamed model, a cytokine cocktail containing IL-1β (25 ng/mL) and TNF-α (50 ng/mL) was added to the basolateral (BL) once daily for 72 h. Finally, cell culture media from both apical (AP) and BL compartments were collected from the inflamed cells for IL-8 quantification. mRNA and protein were extracted from cells in both the standard and inflamed models and analysed through ddPCR and ELISA, respectively. A comparison of ( B ) IL-8 secretion and ( C ) SGLT1, ( D ) GLUT2, ( E ) ACE2 and ( F ) TMPRSS2 mRNA expression between standard (grey) and inflamed (blue) Caco-2/TC7 cells is shown. Total RNA was extracted and reverse transcribed, and absolute copies of SGLT1, GLUT2, ACE2 or TMPRSS2 cDNA measured by ddPCR are expressed relative to the housekeeping gene TBP. For IL-8 detection, cell culture media were processed, and IL-8 measured using ELISA and corrected for total protein measured by Bradford assay. Data are presented as mean ± SD (n = 6 technical replicates from N = 3 biological replicates). Differences between cells in the standard and inflamed environments in B-F were determined by unpaired t -test with Welch’s correction; ns—not significant, ** P < 0.01, *** P < 0.001. A —created with BioRender.com ( https://BioRender.com/r16s632 ); B-F —created with GraphPad Prism version 9.0.1.

Article Snippet: High Capacity RNA-to-cDNA reverse transcription kits, and FAM- or VIC-labelled TaqMan primers for ACE2 (Hs01085333_m1), SGLT1 (Hs01573793_m1), GLUT2 (Hs01096908_m1), TMPRSS2 (Hs01122322_m1) and TBP (Hs00427620_m1) were also from Thermo Fisher Scientific.

Techniques: Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison, Expressing, Reverse Transcription, Bradford Assay

Journal: Scientific Reports

Article Title: Dexamethasone enhances intestinal glucose absorption and TMPRSS2 expression with implications for hyperglycaemia and infection risk

doi: 10.1038/s41598-025-22312-8

Figure Lengend Snippet: Gene expression changes in dexamethasone-treated and untreated normal and inflamed enterocytes from wild-type C57BL/6 J mice. ( A ) A summary of the GSE113691 study and the samples considered for analysis. Raw counts obtained from the NCBI GEO database ( GSE113691 ) were filtered (< 1 count per million in at least 3 samples) and were normalised using the trimmed mean of m-values (TMM). Differences in ( B ) SGLT1, ( C ) TMPRSS2, ( D ) GLUT2, ( E ) ACE2 gene expression in enterocytes from normal mice (NM) and inflamed mice (IF) between dexamethasone-treated (NM-D, IF-D) and untreated mice (NM-C, IF-C). Volcano plot showing ( F ) 910 differentially expressed genes between NM-D and NM-C and ( I ) 386 differentially expressed genes between IF-D and IF-C. Venn diagrams depicting upregulated and downregulated genes in ( G ) NM-D and ( J ) IF-D treatment groups compared to controls NM-C and IF-C, respectively. Red indicates upregulation and blue indicates downregulation (log 2 FC ≥ 1, FDR ≤ 0.05), overlapping indicates the unchanged number of genes. ( H ) Log 2 FC of SGLT1, TMPRSS2, GLUT2, and ACE2 between NM-D vs. NM-C and IF-D vs. IF-C (*indicates significantly differentially expressed genes (log 2 FC ≥ 1, FDR ≤ 0.05)). Gene Set Enrichment Analysis on the mouse Hallmark signature collection, showing positively and negatively enriched gene sets in ( K ) NM-D vs. NM-C and ( L ) IF-D vs. IF-C. Orange bars indicate gene sets enriched in both groups. ( M ) Pearson correlation matrix showing associations between SGLT1, TMPRSS2, GLUT2, and ACE2 mRNA expression in mouse intestinal epithelial cells. Blue represents positive correlation, red represents negative correlation. ( B-E ) Data are presented as mean ± SD (N = 3). ( B-E ) Significant differences were determined using one-way ANOVA with Fisher’s LSD multiple comparisons test. ns—not significant, * P < 0.05, ** P < 0.01, *** P < 0.001. DEX—dexamethasone, NM-C—normal mouse control, NM-D—normal mouse treated with dexamethasone, IF-C—inflamed mouse control, IF-D—inflamed mouse treated with dexamethasone, NES—normalised enrichment score.

Article Snippet: High Capacity RNA-to-cDNA reverse transcription kits, and FAM- or VIC-labelled TaqMan primers for ACE2 (Hs01085333_m1), SGLT1 (Hs01573793_m1), GLUT2 (Hs01096908_m1), TMPRSS2 (Hs01122322_m1) and TBP (Hs00427620_m1) were also from Thermo Fisher Scientific.

Techniques: Gene Expression, Expressing, Control

Journal: Scientific Reports

Article Title: Dexamethasone enhances intestinal glucose absorption and TMPRSS2 expression with implications for hyperglycaemia and infection risk

doi: 10.1038/s41598-025-22312-8

Figure Lengend Snippet: Dexamethasone downregulates GLUT2 mRNA in Caco-2/TC7 cells. The acute and chronic effect of dexamethasone (DEX) on GLUT2 mRNA expression was assessed in 14-day differentiated Caco-2/TC7 cells, under either standard conditions (Panel 1) or an inflamed state induced by daily exposure to IL-1β and TNF-α during the final 72 h (Panel 2). Cells were treated with dexamethasone (5, 10, 20 µM) or 0.1% (v/v) DMSO vehicle control for different time durations: ( A, D ) 4 h, ( B, E ) 24 h and ( C, F ) 60 h. A comparison of the effect of dexamethasone between standard (grey) and inflamed (blue) conditions is shown in Panel 3 ( G —4 h, H —24 h, I —60 h). Total RNA was extracted and reverse transcribed, and absolute copies of GLUT2 cDNA measured by ddPCR are expressed relative to the housekeeping gene TBP. Values represent mean fold change compared to DMSO vehicle control (Panel 1) or cytokine-treated DMSO vehicle control (Panel 2). Data are presented as mean ± SD (n = 6 technical replicates from N = 3 biological replicates). Differences were determined as follows: A, D, G —unpaired t -test with Welch’s correction; B-C, E–F —one-way ANOVA with Fisher’s LSD multiple comparisons test; H, I —two-way ANOVA with Fisher’s LSD multiple comparisons test. ns—not significant, * P < 0.05, ** P < 0.01, *** P < 0.0001. Graphs created with GraphPad Prism version 9.0.1.

Article Snippet: High Capacity RNA-to-cDNA reverse transcription kits, and FAM- or VIC-labelled TaqMan primers for ACE2 (Hs01085333_m1), SGLT1 (Hs01573793_m1), GLUT2 (Hs01096908_m1), TMPRSS2 (Hs01122322_m1) and TBP (Hs00427620_m1) were also from Thermo Fisher Scientific.

Techniques: Expressing, Control, Comparison, Reverse Transcription

Journal: Scientific Reports

Article Title: Dexamethasone enhances intestinal glucose absorption and TMPRSS2 expression with implications for hyperglycaemia and infection risk

doi: 10.1038/s41598-025-22312-8

Figure Lengend Snippet: Pearson correlation analysis. ( A ) Pearson correlation matrix showing the association between variables. Blue represents a positive correlation, and red represents a negative correlation. ( B ) Table showing the statistical significance of the relationship between variables examined by Pearson correlation analysis. ACE2, TMPRSS2, SGLT1 and GLUT2 are mRNA expression levels; IL-8 is a secreted protein. The correlation matrix was created with GraphPad Prism version 9.0.1. ns—not significant; *** P < 0.001.

Article Snippet: High Capacity RNA-to-cDNA reverse transcription kits, and FAM- or VIC-labelled TaqMan primers for ACE2 (Hs01085333_m1), SGLT1 (Hs01573793_m1), GLUT2 (Hs01096908_m1), TMPRSS2 (Hs01122322_m1) and TBP (Hs00427620_m1) were also from Thermo Fisher Scientific.

Techniques: Expressing

Journal: Nature Communications

Article Title: The Hippo terminal effector YAP boosts enterovirus replication in type 1 diabetes

doi: 10.1038/s41467-025-64508-6

Figure Lengend Snippet: A Scheme how β-YAP mice were generated by crossing RIP-rtTA with TetO-YAP Ser127A mice. B IHC and C Western Blot confirmation of YAP induction in pancreatic islets after 2 days i.p injection of Dox. D YAP was transiently induced by doxycycline (DOX) administration in drinking water for 2 weeks (β-YAP) and results compared to -DOX/-YAP (control; C ). E –H intraperitoneal glucose tolerance test (ipGTT) and respective AUC analyses in β-YAP and control male ( E , F; n = 16 C, n = 18 β-YAP) and female ( G , H , n = 12/group) mice. I , J Insulin levels during an ipGTT measured before (0 min) and 15/30 min after glucose injection in β-YAP and control male ( I; n = 11/group) and female ( J ; n = 9 C, n = 11 β-YAP) mice. K , L Islets were isolated from β-YAP and control mice, cultured overnight and subjected to an in vitro GSIS. K Insulin secretion during 1 h-incubation with 2.8 mM (basal) and 16.7 mM glucose (stimulated), normalized to insulin content and ( L ) stimulatory index denotes the ratio of stimulated to basal insulin secretion ( n = 11). M RT-PCR for MafA, Nkx6.1, Slc2a2, NeuroD1, GCK, Ins1, Ins2, Pdx1, Glis3 ( n = 6 C, n = 11 β-YAP mice), Nkx2.2 ( n = 6 C, n = 10 β-YAP mice), Abcc8, and Kcnj11 (n = 3 C , n = 4 β-YAP mice). Microscopical analyses of β-proliferation by Ki67 ( N , O ) and pHH3 ( P , Q ) in both ( N , P ) male and ( O , Q ) female mice expressed as percentage of Ki67- ( n = 6 mice/group) or pHH3- ( n = 6 male and n = 5 female mice/group) positive β-cells. Insulin-positive area ( R , T ) and β-cell mass ( S , V ) in both ( R , S ) male and ( T , U ) female mice ( n = 6 mice/group). Data are expressed as means ± SEM. P- values were calculated by two-tailed unpaired Student t- test for all except by a mixed-effects model with Holm-Sidak multiple comparisons correction for ( E , G ). *** p < 0.001 compared to control; Scale bars depict 20 µm. Source data are provided as a Source Data file.

Article Snippet: TaqMan® Gene Expression Assays were used for Stk4 (#Hs00178979), CTGF (#Hs01026927-g1), CXCL10 (#Hs00171042), IFNB1 (#Hs02621180), OAS1 (#Hs00973637), DDX58 (#Hs01061436), TLR3 (#Hs01551078), IFIH1 (#Hs00223420), IL6 (#Hs99999032), YAP1 (#Hs00371735), AMOTL2 (Hs#01048101), ANKRD1 (Hs#00173317), Tuba1a (#Hs00362387), ACTB (#Hs99999903), Stk4 (#Mm00451755), Tuba1a (#Mm00846967), Stk4 (#Rn01750112), ACTB (#Rn00667869), Nkx2.2 (#Mm00839794_m1), Nkx6.1 (#Mm00454961_m1), NeuroD1 (#Mm01946604_s1), MafA (#Mm00845206_s1), Slc2A2 (#Mm00446229_m1), Abcc8 (#Mm00803450_m1), Glis3 (#Mm00615386_m1), Gck (#Mm00439129_m1C41), KcnJ11 (#Mm00440050_s1), Ins1 (#Mm04207513_g1), Ins2 (#Mm00731595_g1), PDX1 (#Mm00435565_m1), and), and ACT (#Mm00607939_s1).

Techniques: Generated, Western Blot, Injection, Control, Isolation, Cell Culture, In Vitro, Incubation, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test