Journal: Aging Cell

Article Title: Reduced Mitochondrial Adenine Nucleotide Translocase 1 ( ANT1 ) Correlates With Aging‐Associated Airway Remodeling

doi: 10.1111/acel.70264

Figure Lengend Snippet: SLC25A4 (ANT1) expression in human lungs is decreased in an age‐associated manner. (A–C) Single cell RNA sequencing from a healthy human dataset from the Human Lung Cell Atlas shows that SLC25A4 expression (log normalized, ln) decreases with age in the (A) airway basal cells ( r = −0.5151, p < 0.0001), (B) ciliated bronchial epithelial cells ( r = −0.3246, p < 0.0001), and (C) AT2 cells ( r = −0.1214, p < 0.0001). Tables in the data show the number of total cells analyzed (samples), number of human donors, number, and percentage of cells with expression of SLC25A4 (ANT1). Statistics for panels A–C by Pearson's r test with p ‐values in the legend. (D) Bulk RNA sequencing from the Tabula Muris database shows decreased Slc25a4 expression in female and male mice lungs over age. (E) Western blot and quantification relative to total protein intensity for Ant2 from mouse lungs of wildtype and Ant1 knockout (A1KO) mice, young and old ( n = 3–4 mice per group). * p < 0.05. Statistics by 2‐way ANOVA with Tukey's post‐test. Data are shown as mean ± SEM.

Article Snippet: Bulk RNA sequencing data from a mouse aging cell atlas database (Tabula Muris) showed decreased Slc25a4 expression in the lungs of female mice with increased age after 9 months old and a reduction in male mice after 18 months old (Figure ).

Techniques: Expressing, RNA Sequencing, Western Blot, Knock-Out

Journal: Aging Cell

Article Title: Reduced Mitochondrial Adenine Nucleotide Translocase 1 ( ANT1 ) Correlates With Aging‐Associated Airway Remodeling

doi: 10.1111/acel.70264

Figure Lengend Snippet: Loss of ANT1 resulted in dysregulation of collagen 8 in the airway epithelium with aging. (A) Bulk RNA sequencing was performed on human Beas‐2b cells with siRNA knockdown of ANT1 compared to non‐targeting control siRNA. Differential expression gene (DEG) analysis was performed and the top up and down‐regulated DEGs are shown with adjusted p < 0.05. (B) Key collagen genes that were upregulated or downregulated at a p < 0.05. (C) Western blot analysis of COL8A1 expression in Beas‐2b cells with Crispr‐Cas9 knockout of ANT1 compared to scrambled sgRNA control cells. Results were normalized to total protein intensity per lane. Statistics were calculated using unpaired Student's t ‐test. ** p < 0.001. (D) Immunofluorescence for Col8a1 in the CC10+ airway cells of mouse lung tissue sections. Col8a1 is shown in grayscale and composite with transmitted light brightfield, N = 4 for old WT and n = 3 for old A1KO. Data are shown as mean ± SEM. Statistics by Two‐way ANOVA with Tukey's post‐test. * p ‐value noted. (E) Immunofluorescence for COL8A1 in the ciliated cells (Acetyl‐α‐tubulin+) of the airways of healthy human lungs from young (31–45 years old, n = 6, 2 male and 4 females) versus aged subjects (73–76 years old, n = 6, 3 males and 3 females) separated by age range. Immunofluorescence data is reported as Integrated Density values from ImageJ. Data are shown as mean ± SEM. * p ‐value noted. Statistics were calculated using unpaired Student's t ‐test with Mann–Whitney post‐test.

Article Snippet: Bulk RNA sequencing data from a mouse aging cell atlas database (Tabula Muris) showed decreased Slc25a4 expression in the lungs of female mice with increased age after 9 months old and a reduction in male mice after 18 months old (Figure ).

Techniques: RNA Sequencing, Knockdown, Control, Quantitative Proteomics, Western Blot, Expressing, CRISPR, Knock-Out, Immunofluorescence, MANN-WHITNEY