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. 2021 Jul 15;13(8):e13610. doi: 10.15252/emmm.202013610

Figure EV2. WWOX‐KO cerebral organoids showed enhanced astrogenesis and loss of checkpoint inhibition.

Figure EV2

  1. Western blot analysis of week 24 COs, examining the protein expression of WWOX, GFAP (astrocytes), SOX2 (RG cells), and NeuN (mature neurons).
  2. A summary of the band intensities of the Western blot exemplified in (A), presented as mean ± SEM of the WT COs and KO COs, and as fold change compared with the WT COs (WT: n = 2 from 1 batch; KO: n = 4 from 1 batch).
  3. IF staining of week 10 COs with the proliferation marker Ki67 localized with S100β (astrocytes) and SOX2 (RGs) (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches).
  4. Quantification of (C). The boxplot represents the 1st and 3rd quartile, with its whiskers showing the minimum and maximum points and a central band representing the median. Statistical significance was determined using the two‐tailed unpaired Welch’s t‐test (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches).
  5. Week 10 COs stained for Ki67 (proliferation), γH2AX (DNA breaks) and SOX2 (RGs) (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches).
  6. Quantification of γH2AX+/Ki67+ double‐positive cells (DPCs) normalized to the number of SOX2+ cells, corresponding to the size of the VZ, seen in (C). The boxplot represents the 1st and 3rd quartile, with its whiskers showing the minimum and maximum points and a central band representing the median. Statistical significance was determined using the two‐tailed unpaired Welch’s t‐test (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches).
  7. Week 10 COs co‐stained for the apoptosis marker cleaved caspase‐3 and the RG marker SOX2 in the VZ of WT, KO and W‐AAV COs (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches; and W‐AAV: n = 4 from 1 batch). Scale = 50 µm.
  8. Quantification of the data presented in (G). The boxplot represents the 1st and 3rd quartile, with its whiskers showing the minimum and maximum points and a central band representing the median. Statistical significance was determined using one‐way ANOVA with Tukey’s multiple comparisons test (WT: n = 6 from 2 batches; KO: n = 4 from 2 batches; W‐AAV: n = 4 from 1 batch).

Data information: n.s (non‐significant), *P ≤ 0.05, **P ≤ 0.01.