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. 2022 Nov 7;13(11):934. doi: 10.1038/s41419-022-05370-y

Fig. 5. Rab10 helps surface expression of EGFR in macrophages.

Fig. 5

A EGFR is localized to Rab10-positive Golgi and early endosomes compartments. RAW2654.7 cells were immunostained with anti-TGN38K (red), anti-EEA1 (red), or anti-LAMP1 (red) and anti-Rab10 (green) Abs. Representative confocal images show colocalization (yellow) of Rab10 and TGN38K, EEA1, LAMP1. Scale bar, 5 μm. B Rab10 colocalized with EGFR in RAW264.7 cells. Scale bar, 5 μm. C Immunoblot analysis of EGFR immunoprecipitates of lysates of RAW264.7 cells. D Immunoblot analysis of Rab10 immunoprecipitates of lysates of RAW264.7 cells. E–G Sh-ctrl and sh-Rab10 BMDM were treated with LPS (1 μg/mL) for 12 h. E EGFR expression on the surface of BMDM was analyzed by flow cytometry. F Percentage of EGFR-positive BMDM is shown (n = 3). G Mean fluorescence intensity (MFI) is shown (n = 3). H Stable expression of Rab10-WT, Rab10-T23N or Rab10-Q68L mutant in Rab10 shRNA knockdown RAW264.7 cells. Immunoblotting of whole-cell extracts from the indicated cells using a-Rab10 or a-GAPDH as a loading control. I Immune-staining of TGN38 and indicated mutations of Rab10 in RAW264.7. J Immune-staining of EEA1 and indicated mutations of Rab10 in RAW264.7. K–M The indicated Rab10 mutant RAW264.7 cells were stimulated with LPS for 12 h. K EGFR expression on the surface of BMDM was analyzed by flow cytometry (n = 3). L Percentage of EGFR-positive BMDM is shown (n = 3). M Mean fluorescence intensity (MFI) is shown (n = 3). The graphs depict mean ± SD based on three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.