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. 2022 Jan 28;70(5):989–1004. doi: 10.1002/glia.24153

FIGURE 3.

FIGURE 3

Changes in inflammatory and neurotrophic gene expression after astrocyte GATA6 manipulation. (a) Confirmation of GATA6 transcript reduction (SMA) or overexpression (HC) after exposure to GFP control, GATA6 expressing, or GATA6KD lentivirus and (b) associated change in NFĸB transcript production. (c, d) representative images of GATA6 and phNFĸB protein expression changes after lentiviral manipulation assessed by ICC ([c]: GATA6 = red, S100β = white, DAPI = blue; [d] phNFĸB = pink, NFĸB = teal, DAPI = blue) ([e, f] quantification). (g) Secreted IL6 protein levels confirm transcript changes after GATA6 manipulation. (h) Significant decrease in IL6, C1q, and C3 transcripts after GATA6KD in SMA astrocytes, trends for increases after GATA6OE in HC astrocytes. (i) no significant changes in neurotrophic support after GATA6 manipulation. (j) Significant decrease in reactive GFAP transcripts, but importantly not in characteristic marker S100β. ICC quantification displays relative fluorescence (total fluorescence for each channel normalized to DAPI) for each image to account for differences in cell density between ROIs. (2‐way ANOVA, Tukey's multiple comparisons test for qPCR; 1‐way ANOVA, Tukey's multiple comparisons test for ICC, *p value<0.05, ** < 0.005, *** < 0.0005, **** < 0.0001)