Journal: Neuroglia (Basel, Switzerland)
Article Title: Aging Effects on Metabolic Sensor and Glycogen Metabolism in Old Male versus Female Rat Primary Hypothalamic Astrocyte Cultures
doi: 10.3390/neuroglia6040041
Figure Lengend Snippet: Effects of Glucose Transporter-2 (GLUT2) Gene Knockdown on Old Male and Female Hypothalamic Primary Astrocyte GLUT2 Protein Expression. Old male and female rat astrocyte cultures were pretreated with scramble (SCR) or GLUT2 siRNA prior to incubation in 5.5 (G5.5) or 0 (G0) mM glucose-containing media. Young adult male or female SCR siRNA/G5.5 astrocyte cultures served as controls. Astrocyte lysates were analyzed across treatment groups by Western blot for GLUT2 protein content in three independent experiments. Target protein optical density (O.D.) measures acquired in a Bio-Rad ChemiDoc ™ Touch Imaging System were normalized to total in-lane protein (loading control) using Stain-Free technology and Bio-Rad Image Lab ™ 6.0.0 software. Data depict mean normalized GLUT2 protein O.D. values ± S.E.M. for male and female astrocyte treatment groups. In each figure, the solid bar at left depicts mean GLUT2 O.D. for young adult astrocyte SCR siRNA/G5.5 cultures, whereas old male or female astrocyte treatment groups are illustrated as follows: SCR siRNA/G5.5 (horizontal-striped bars); GLUT2 siRNA/G5.5. (diagonal-striped bars); SCR siRNA/G0 (crosshatched bars); GLUT2 siRNA/G0 (vertical-striped bars). For each sex, mean normalized GLUT O.D. data were compared between young and old SCR siRNA/G5.5 groups by t test and old astrocyte treatment groups were analyzed by two-way ANOVA and Student-Newman-Keuls post-hoc , using GraphPad Prism, Vol. 8 software. Statistical differences between treatment group pairs are indicated by the following symbols: * p < 0.05; ** p < 0.01; *** p < 0.001. Tables below and summarize results shown in graphic format; results from prior studies involving SCR siRNA/G5.5, GLUT2 siRNA/G5.5, SCR siRNA/G0, or GLUT2 siRNA/G0 treatment on young adult male or female astrocyte GLUT2 protein expression are presented for comparison. Red font denotes a sex difference in age effect on SCR siRNA/G5.5 control group GLUT2 protein expression. Blue font indicates an age-related change in treatment effect on GLUT protein profiles.
Article Snippet: Quantified Western blot target protein optical densities (O.D.) were normalized to total in-lane protein using a Bio-Rad ChemiDoc TM Touch Imaging System and Image Lab TM 6.0.0 software.
Techniques: Knockdown, Expressing, Incubation, Western Blot, Imaging, Control, Staining, Software, Comparison
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