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. 2022 Dec 22;18(1):237–253. doi: 10.1016/j.stemcr.2022.11.018

Figure 3.

Figure 3

Human astrocytes secrete C4

(A) C4 secretion normalized to number of nuclei. Each circle represents the average of eight wells per biological replicate per line; n = 3 independent differentiations for 1016A, Hues8, Mito234, and Mito80, and n = 2 for H1. The lower panel represents the total copy number variation of C4 per line. Ordinary one-way ANOVA, Tukey’s multiple comparisons test. n.s., non-significant.

(B) Normalized C4 secretion measured by ELISA comparing the effects of monensin and IFN-γ in hASTROs. Data represent individual wells (n = 8) of three biological triplicates (squares, circles, and rhombi) per line. Statistics available in Table S1.

(C) Schematic of experimental design to assess C4 detection at synaptosomes using C4 KO and C4 WT-hASTROs and Ngn2 differentiated neurons.

(D) Western blots of C4 expression in C4WT and KO-hASTROs, total lysates, and concentrated astrocyte-conditioned media (ACM) (left panel); C4 expression on synaptosomes purified from C4-WT and KO-Ngn2 neurons incubated with C4 WT or C4 KO ACM in different combinations (right panel). Ponceau and actin are used as loading controls.