Single-cell transcriptomics reveals distinct inflammation-induced microglia signatures

Sousa, Carole; Golebiewska, Anna; Poovathingal, Suresh K; Kaoma, Tony; Pires-Afonso, Yolanda; Martina, Silvia; Coowar, Djalil; Azuaje, Francisco; Skupin, Alexander; Balling, Rudi; Biber, Knut; Niclou, Simone P; Michelucci, Alessandro

We are in agreement with the reviewer's remark, which is in line with Referee #1 comment (please refer to minor point 3b), therefore we performed additional experiments to include a positive control (i.e. bone marrow monocytes). Briefly, as mentioned above, we used the Monocyte Cell Isolation Kit (Miltenyi Biotec) that has been developed for the isolation of monocytes from suspensions of mouse bone marrow. Remarkably, FACS analyses revealed pure populations of Ly6C + cells after depletion of magnetically labelled non-target cells, i.e. Fig. EV2 and the corresponding qPCR data in Fig. 2D , which further support the validity of our FACS gating strategy and purity of FACS-sorted cells. We revised the text and the figure legends accordingly. Figure 3C We thank the reviewer for highlighting these imprecisions. We corrected them accordingly. Please consider that the volcano plot in Fig. 3C is now shown in Appendix Fig. S3 . Thank you for the submission of your revised manuscript to our editorial offices. We have now received the reports from the referees that were asked to re-evaluate your study (you will find enclosed below).