Fig. 4

Transcriptomic changes of KCs after GLY treatment. A UMAP visualization of the five subtypes for KCs. B Heatmap of the expression levels of representative marker genes in KC subtypes. C Histogram showing proportional changes of each KC subtype for two groups. D GO analysis for upregulated DEGs of different cell subtypes. E Boxplots of ROS score for each cell subtype. The middle lines indicate median values, boxes range from the 25th to 75th percentile. P value is from wilcox.test (unpaired and two-tailed); ***P < 0.001, *P < 0.05. F Split violin plots of the expression of four genes associated with phagocytosis. G Violin plots comparing the expression level of Spp1 in each subtype between control and GLY groups (left). GO analysis for upregulated DEGs of KC4 involving Spp1 gene (right). H Typical images and bar graph showing the increased expression level of CD68 after GLY treatment. Boxed regions are amplified on the right; scale bar, 100 μm; error bar, mean ± SD; P value is from Student’s t-test (unpaired and two-tailed); ****P ≤ 0.0001; the quantification was based on six random areas from each group. I Heatmap of upregulated and downregulated genes for KC4 in control and GLY samples. J Differentially expressed cytokines for each KC subtype after GLY treatment. K Heatmap for common upregulated and downregulated genes of all KC clusters