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. 2023 Jun 26;42(15):e111247. doi: 10.15252/embj.2022111247

Figure 6. cADPR/calcium signaling regulates release of SPARCL1 from astrocytes.

Figure 6

  • A
    A schematic drawing cADPR/calcium signaling showing that cADPR produced by the catalytic action of CD38 from NAD+ acts as a calcium releasing second messenger via ryanodine receptors (RyR). 8‐Bromo‐cADPR (8Br), a cADPR antagonist; 2APB, a IP3 inhibitor; and Dantrolene (Dant), a ryanodine receptor inhibitor.
  • B
    Coomassie staining shows the total protein composition of ACM. Representative western blot images of SPARCL1 and β‐actin protein expression in ACM and cell lysates from WT astrocytes treated with 8‐Br‐cADPR (50 μM), 2APB (20 μM), or Dantrolene (20 μM).
  • C, D
    Quantification of SPARCL1 levels in ACM and cell lysates. Protein levels in ACM and lysates were normalized to total protein and the loading control β‐actin, respectively (n = 5 independent culture per condition from five animals, one‐way ANOVA followed by the Tukey–Kramer test).
  • E
    RT‐qPCR analysis for the expression of Ryr2 mRNA in ACSA2 negative or positive cells from the mPFC of ctrlP10 and CD38 AS‐cKOP10 mice. Astrocytes were enriched by magnetic bead assisted cell sorting (MACS) for the astrocytes‐specific ACSA2 antigen (n = 4 animals per genotype, two‐way ANOVA followed by Bonferroni's multiple comparisons test).

Data information: Data represent means ± SEM. *P < 0.05.

Source data are available online for this figure.