Fig. 8

A distinctive M2 type activation in alveolar macrophages after coal exposure A A single-cell sequencing plot shows the effects of coal exposure and Vitamin D supplemented coal exposure on the polarization of macrophages to the M2-type. B The group’s distribution of canonical cell markers CD86, Mrc1, and MLPH target gene transcripts across the t-distributed stochastic neighbor embedding (tSNE) plot. The subset of M1 cells was identified by CD86 expression, and the M2 subset was identified by Mrc1 expression. The color key indicates MAGIC-imputed gene expression values. C Violin plots display the distribution of the expression of previously reported M2 polarization-related markers (Mrc1, up), M1 polarization-related markers CD86, bottom). D Double immunofluorescence staining of MLPH and CD206 verified M2-type macrophages that emerged after coal exposure and disappeared after the Vitamin D treatment in the lung tissue and Raw267.4 cells both in vivo and in vitro. Scale bar, 40 μm. E The RAW264.7 cells were treated with phosphate-buffered saline (PBS; control), coal dust, and a combination of coal dust and Vitamin D; the whole-cell lysates were harvested 24 h after the treatment. Western blots determined the expression level of MLPH. β-actin was used as the internal control