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. Author manuscript; available in PMC: 2023 Sep 1.
Published in final edited form as: Neuroglia. 2023 Jun 21;4(3):158–171. doi: 10.3390/neuroglia4030011

Figure 4. GLUT2 Gene Silencing Effects on Male and Female Hypothalamic Primary Astrocyte Total and Phosphorylated 54/46 kDa SAPK/JNK Protein Expression.

Figure 4.

Pooled astrocyte lysates were measured in triplicate using Bio-Rad Stain-Free Imaging equipment and software. Protein optical density (O.D.) values obtained in our Bio-Rad ChemiDoc™ Touch Imaging System were related, Bio-Rad Image Lab™ 6.0.0 software, to total in-lane protein, e.g. all protein electrophoresed in the individual sample lane, for normalization. Data depict mean normalized total (Figures 4A and 4C) or phosphorylated (Figures 4B and 4D) 54 and 46 kDa SAPK/JNK protein O.D. values ± S.E.M. for male (M; gray bars, at left) and female (F; white bars, at right) G5.5- or G0-exposed astrocytes pretreated with SCR versus GLUT2 siRNA. Experimental groups consisted of male or female rat astrocyte cultures that were exposed to the following treatments: G5.5/SCR siRNA [solid bars (gray, M; white, F)]; G5.5/GLUT2 siRNA [horizontal-stripe bars (gray, M; white, F)]; G0/SCR siRNA [diagonal-stripe bars (gray, M; white, F)]; or G0/GLUT2 siRNA [cross-hatch bars (gray, M; white, F)]. GraphPad Prism (Volume 8) software was used to analyze averaged normalized protein O.D. values by three-way ANOVA, followed by Student-Newman-Keuls post-hoc test; results are presented in Supplementary Table 1. Open circles depict individual independent data points. Statistical differences between discrete pairs of treatment groups are denoted as follows: *p < 0.05; **p < 0.01; ***p < 0.001.