Extended Data Fig. 8: Ex vivo CCR2 silencing and in vivo biological effects in aNP-treated MC38 tumour-bearing mice.
a. Screening of siCCR2 constructs (Supplementary Table 4) using bone marrow-derived macrophages. Transfection was performed with aNP18-siCCR2 or aNP18-siCtrl, at concentrations 1, 10, and 100 nM siRNA, respectively. Expression levels of Ccr2 were determined by RT-qPCR and the ΔΔCq-values are displayed. The primers are listed in Supplementary Table 3. Data represents mean ± SD of one experiment (minimal of n = 2 biological replicates). b. Number of myeloid cells per gram of tumour tissue as determined by flow cytometry. Data represent mean ± SD of one biological replicate (n = 4-5 mice) and analyzed by student’s t-test c. Fraction of CCR2-positive myeloid cells within tumour tissue expressed as percentage (%) of myeloid cells. Data represent mean ± SD of one experiment (n = 4-5 mice) and analyzed by student’s t-test. Statistically significant differences between aNP18-siCtrl versus aNP18-siCCR2 formulations are indicated by: ** indicates statistical significance (p = 0.0084). d. Individual tumour volumes of MC38 tumour-bearing mice (n = 5 per group) receiving repeated intravenous administrations of aNP18 containing scrambled siRNA (aNP18-siCtrl) or CCR2 siRNA (aNP18-siCCR2) (0.5 mg/kg per administration, cumulative dose 3.5 mg/kg). e. Individual mouse body weights (n = 5 mice).
