Extended Data Fig. 7: The radioresistant compartment is dispensable for autoinflammation in APLAID.
a) RT-qPCR in FACS sorted immune cells from skin (left) and non-immune cells (right) from lung tissue in PLCγ2S707Y/+ compared to PLCγ2+/+ controls reveals elevated G-CSF transcripts in both skin myeloid cells (monocytes, macrophages, Langerhans Cells) and fibroblasts. Two independent experiments from 4 different mice per genotype are shown. Animals were 4–6 weeks old. b) Scheme of BM chimera generation (in dark yellow). PLCγ2S707/+ (in yellow) are the donor for lethally irradiated PLCγ2S707Y/+ recipients (yellow). Single cell suspension of BM cells (1 × 106/mL) are transplanted by i.v. injection into recipient animals 3 hrs after irradiation. c) Severity of skin inflammation is determined by the APLAID skin score on a 0–5 scale post weaning (n = 3 mice per group). d) A growth curve exhibits a continuous weight loss in PLCγ2S707Y/+ into PLCγ2S707Y/+ chimeras (n = 3 mice per group). e) Splenomegaly persists in the PLCγ2S707Y/+ into PLCγ2S707Y/+ chimera (n = 3 mice per group). f) Kaplan-Meier analysis demonstrate decreased survival rates in PLCγ2S707Y/+ into PLCγ2S707Y/+ chimera (n = 3 mice per group). g-j) ADVIA analyzer data of the blood reveal increased neutrophil and decreased lymphocyte counts in PLCγ2S707Y/+ receiving BM from PLCγ2S707Y/+ mice, whilst thrombo- and erythropoiesis remain unaltered (n = 3 mice per group). k) Proportion between circulating CD45.1+ immune cells from the WT donor (6–8 weeks of age) and CD45.2+ radioresistant immune cells from the recipient (4 weeks of age) 7 weeks post-transplantation (n = 3 mice per group). PLCγ2S707Y/+ mice were 4-week-old. Error bars represent mean ± SEM. Statistical significance for skin score was determined by a two-way ANOVA. Statistical significance for longitudinal weight data was determined by a paired two-sided t-test. Spleen weights between two groups were determined by an unpaired two-sided Student’s t-test. Statistical significance for the survival curve was determined by a Mantel-Cox test. G-CSF levels and cell numbers between two groups were assessed by an unpaired two-sided Student’s t-test.
