Fig. 2: In vivo engulfment of different fluorescent proteins following optic nerve crush. | Nature Communications

Fig. 2: In vivo engulfment of different fluorescent proteins following optic nerve crush.

From: FEAST: A flow cytometry-based toolkit for interrogating microglial engulfment of synaptic and myelin proteins

Fig. 2

a pKa values for fluorescent proteins (ZsGreen, TdTomato, EGFP, and EYFP) used to label retinal ganglion cells by crossing CHX-10-cre mice with lox-stop-lox (lsl) reporter mice. b MFI of ZsGreen contained in microglia from CHX10cre+lsl:ZsGreen mice. n = 5 (2 females and 3 males) for ONC and n = 4 (4 males) for No crush. c MFI of TdTomato contained in microglia from CHX10cre+lsl:TdTomato mice. n = 8 for ONC (2 females and 6 males) and n = 5 (5 females) for No crush. d MFI of EGFP contained in microglia from CHX10cre+lsl:EGFP mice. n = 5 for ONC (4 females and 1 male) and n = 5 for No crush (4 females and 1 male). e MFI of engulfed EYFP contained in microglia from CHX10cre+lsl:EYFP mice. n = 5 (5 males) for ONC and n = 4 (2 females and 2 males) for No crush. b–e Microglia were isolated from LGNs/SCs by Dounce homogenization at 4 °C and enriched by Percoll centrifugation 3 days after bilateral ONC. Microglia are gated on live, single cells, CD45+, CD11b+, CX3CR1+, Ly-6C- or CD206-. MFI values are normalized to the mean MFI of the No crush controls. The gates for positive microglia on the flow cytometric plots (5% contour plots) are based on the fluorescent signal from WT cortical microglia (<1% cells in positive gate) for each experiment. Error bars indicate standard error of the mean. Statistical analysis: unpaired two-tailed t test. Source data provided.

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