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. 2014 Dec 15;127(24):5228–5239. doi: 10.1242/jcs.154393

Fig. 4.

Fig. 4.

Endocytosis of the CSF-1R is sensitive to clathrin inhibition. (A) Surface staining of CSF-1R in resting and CSF-1-stimulated BMMs that were transfected once (CHC KD+) or twice (CHC KD++) with RNAi oligos specific to CHC. See Materials and Methods. Scale bar: 5 µm. (B) Quantification of CSF-1R surface staining on resting and CHC-knockdown (KD) BMMs. Norm., normalized. Data show the mean±s.d. (n = 25 cells per condition); *P<0.05 (two-way ANOVA followed by Sidak's multiple comparison test; no difference for CHC KD++). (C) Transferrin uptake in CHC knockdown BMMs. Transferrin was used at 10 µg/ml. Scale bar: 5 µm. (D) Western blot showing reduction of CHC protein levels in CHC KD++ BMM cells. (E) Surface staining CSF1-R on BMMs treated with the CME inhibitory drugs dynasore or PitStop2 and exposed to CSF-1 for the indicated times. Scale bar: 5 µm. (F) Images of fluorescent transferrin uptake in BMMs treated with dynasore or PitStop2 as in C. Scale bar: 5 µm.