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. 2021 Nov 15;12:6565. doi: 10.1038/s41467-021-26851-2

Fig. 8. Genetic interaction of PQBP1 with LRP1 or TREM2 in microglia.

Fig. 8

a Immunocytochemistry revealed high efficiency of tamoxifen-induced Pqbp1-cKO and D10A-mutant Cas9 nuclease/sgRNA-mediated KO of Lrp1/Trem2. Right graphs show quantitative analyses of signal intensities of target genes. Non-TF non-transfected. N = 5 wells. In the analysis of CRIPER-GFP signals: P = 7.35e−12 (Ctrl vs Non-TF), 2.22e−12 (Lrp1-KO vs Non-TF), 3.31e−13 (Trem2-KO vs Non-TF); in the analysis of LRP1 signal: P = 1.09e−11 (Lrp1-KO vs Non-TF); in the analysis of PQBP1 signals: P = 1.92e−14 (Pqbp1-cKO vs Non-TF); in the analysis of TREM2 signals: P = 6.76e−14 (Trem2-KO vs Non-TF); ##P < 0.01 in Tukey’s HSD test (vs Non-TF). b Protocol for double KO of Pqbp1-Lrp1 or Pqbp1-Trem2 genes in primary microglia and their activation of NFκB by Tau 410. Tamox tamoxifen. c RFP signal assay to monitor activation of NFκB by adding Tau 410 in single KO microglia of Lrp1 or Trem2 or double KO microglia of Pqbp1-Lrp1 or Pqbp1-Trem2. N = 5 wells. In tau signal assay of non-induced Pqbp1-cKO group: P = 1.002e−13 [Non-TF (Tau+) vs (Tau−)], 1.005e−13 [Ctrl (Tau+) vs (Tau−)], 5.65e−11 [Lrp1-KO (Tau+) vs (Tau−)], 4.597e−12 [Trem2-KO (Tau+) vs (Tau−)]; in tau signal assay of induced Pqbp1-cKO group: P = 1.005e−13 [Non-TF (Tau+) vs (Tau−)], 1.006e−13 [Ctrl (Tau+) vs (Tau−)], 1.73e−11 [Lrp1-KO (Tau+) vs (Tau−)], 1.98e−11 [Trem2-KO (Tau+) vs (Tau−)]; in RFP signal assay of non-induced Pqbp1-cKO group: P = 4.60e−13 [Non-TF (Tau + ) vs (Tau−)], 1.23e−13 [Ctrl (Tau+) vs (Tau−)], 4.93e−7 [Lrp1-KO (Tau+) vs (Tau−)], 1.15e−12 [Trem2-KO (Tau+) vs (Tau−)]; in RFP signal assay of induced Pqbp1-cKO group: P = 1.005e−13 [Non-TF (Tau+) vs (Tau−)], 1.006e−13 [Ctrl (Tau+) vs (Tau−)], 1.73e−11 [Lrp1-KO (Tau+) vs (Tau−)], 0.9998 [Trem2-KO (Tau+) vs (Tau−)]; #P < 0.05, ##P < 0.01 in Tukey’s HSD test. d Luciferase assay to monitor activation of NFκB by adding Tau 410 to single KO microglia of Lrp1 or Trem2 or double KO microglia of Pqbp1-Lrp1 or Pqbp1-Trem2. N = 4 wells. In non-induced Pqbp1-cKO group: P = 2.20e−11 [Non-TF (Tau+) vs (Tau−)], 3.31e−11 [Ctrl (Tau+) vs (Tau−)], 7.40e−5 [Lrp1-KO (Tau+) vs (Tau−)], 5.76e−15 [Trem2-KO (Tau+) vs (Tau−)]; in induced Pqbp1-cKO group: P = 8.13e−5 [Non-TF (Tau+) vs (Tau−)], 0.0013 [Ctrl (Tau+) vs (Tau−)], 0.0028 [Lrp1-KO (Tau+) vs (Tau−)], 0.9498 [Trem2-KO (Tau+) vs (Tau−)]; #P < 0.05, ##P < 0.01 in Tukey’s HSD test. e A hypothetical scheme of two pathways from extrinsic tau to activation of nuclear NFκB in microglia. Box plots show the median, quartiles, and whiskers that represent data outside the 25th to 75th percentile range.