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. Author manuscript; available in PMC: 2021 Aug 5.
Published in final edited form as: Neuron. 2020 Jun 1;107(3):436–453.e12. doi: 10.1016/j.neuron.2020.05.014

Figure 2: Immunofluorescence and transcription profiling of CD49f+ hiPSC-derived astrocytes confirm expression of canonical markers.

Figure 2:

See also Figure S3.

a) Immunofluorescence images of CD49f+ astrocytes showing expression of astrocyte markers in red or white, and total DAPI cells (cyan). Scale bar, 200μm.

b) Immunofluorescence images of CD49f+ astrocytes showing CD49f+ (cyan), GFAP+ (red), AQP4+ (green) and total DAPI cells (blue). White arrows indicate cells that are CD49f+/AQP4+/GFAP−. Scale bar, 50μm.

c) Percentages of CD49f+ cells across different CD49f+ hiPSC-astrocyte lines that are also GFAP+, AQP4+, and triple positive for GFAP, AQP4, and CD49f. Colored dots correspond to 3 different lines. Error bars show mean ± standard deviation (n=5, 1-2 technical replicates per line).

d) RNA-Seq expression levels of immature astrocyte markers in human CD49f+ astrocytes, expressed in transcripts per million (TPM). Colored dots correspond to 3 different lines. Error bars show mean ± standard deviation (n=9, 3 replicates per line).

e) mRNA expression levels of mature astrocyte markers in human CD49f+ astrocytes, expressed in transcripts per million (TPM). Colors and error bars are as in d).

f) Hierarchical clustering of RNA-Seq data shows that CD49f+ hiPSC-astrocytes (black) closely resemble primary and iPSC-derived astrocytes from independent studies and are distinct from other brain cell types (GEO: GSE73721 in blue; GEO: GSE97904 in pink). Analysis is based on transcriptome-wide expression. CD49f+ samples consist of 3 different lines in 3 replicates each.