Online Repository Figure Legends
Figure E1. A,B, Gene-by-gene view of expression changes of reported PPP-increased genes (A) and PPP-decreased genes (B) in our study. C,D, Gene-by-gene view of expression changes of same number of randomly selected genes. E, F, Expression changes of the group of reported PPP-increased (E) or –decreased (F) genes, compared to randomly selected genes, in our study. Mann-Whitney U test.
Figure E2. Examples of Twist-binding motifs (purple boxes) on examples of PPP - specific genes, including CHST11, SFT2D3, MFGEB, PDGFD, OLFM1, GSTA3, and CYTIP.
Figure E3. mRNA levels of STEAP1 and STEAP4 in various cell types.
Figure E4. Expression of STEAP1 (left) and STEAP4 (right) in different groups of patients in GPP, PPP, and AGEP. Each data point represents the sample from one individual. The center lines show the mean of the different samples in the specified group. *, p<0.05. Mann-Whitney test.
Figure E5. Top signaling pathways enriched in group 3 patients compared to group 2 and group 1 patients, respectively. GPP is shown in black, PPP in green, and AGEP in red.
Figure E6. A) Venn-diagram of genes differentially expressed between the 3 patient groups in GPP, PPP, and AGEP. B) Venn-diagram of checkpoint genes in PPP and GPP. C) Checkpoint genes differentially expressed in PPP, with connectors showing gene interaction based on Genomatix literature search. Red shows genes common to PPP and GPP. D) Checkpoint genes differentially expressed in GPP, with connectors showing gene interaction based on Genomatix literature search. Red shows genes common to PPP and GPP.
Figure E7. Correlation between STEAP expression levels with the those of MEK1/2 targets FOSL1, HIF1A, and CRABP2 in GPP, PPP, and AGEP.Expression values were fitted by linear regression, and slope was tested for significant deviation from 0.
Figure E8. mRNA levels of STEAP1 (white) and STEAP4 (blue) upon scrambled RNAi, STEAP1 RNAi, or STEAP4 RNAi by qRT-PCR. Each bar shows the mean from three experiments. Error bars are the standard error of the mean from the three experiments.
Figure E9. Efficiency of STEAP1 (A) and STEAP4 (B) knockdown under cytokine (TNFa, IL1b, IL36a, IL17a, IL22) treatments, shown by fold change of STEAP1 and STEAP4 mRNA with their respective RNAi compared to scrambled RNAi by qRT-PCR. Each bar shows the mean from three experiments. Error bars are the standard error of the mean from the three experiments.
Figure E10. Cytokine responses independent of STEAP1 and STEAP4. qRT-PCR analyses of the expression levels of various cytokines under the indicated stimulations. Each panel shows 6 conditions: untreated, scrambled RNAi; untreated, STEAP1 RNAi; untreated, STEAP4 RNAi; stimulated, scrambled RNAi; stimulated, STEAP1 RNAi; stimulated, STEAP4 RNAi. Each bar shows the mean from three experiments. Error bars are the standard error of the mean from the three experiments. *, p< 0.05. Student’s t-test.
Figure E11.Cytokine responses independent of STEAP1 and STEAP4. qRT-PCR analyses of the expression levels of various cytokines under the indicated stimulations. Each panel shows 6 conditions: untreated, scrambled RNAi; untreated, STEAP1 RNAi; untreated, STEAP4 RNAi; stimulated, scrambled RNAi; stimulated, STEAP1 RNAi; stimulated, STEAP4 RNAi. Each bar shows the mean from three experiments. Error bars are the standard error of the mean from the three experiments.
Figure E12. ELISA analyses of secreted cytokine levels with scrambled RNAi (white) or STEAP1 RNAi (grey). n=3. Each bar shows the mean from three experiments. Error bars are the standard error of the mean from the three experiments. *, p< 0.05. Student’s t-test.
Figure E13. Overview of the roles of STEAP1 and STEAP4 in inflammatory responses. A, B) Requirement of STEAP1 (A) and STEAP4 (B) in the induction of IL1A, IL1B, IL36G, IL36A, CXCL1 and CXCL8 under various stimuli, summarized from the experiments in Figure 6 and Supplemental Figure 7. Green indicates STEAP is required. C) Schematic of the regulation and function of STEAPs in keratinocytes
Figure E14. Clustering with gene cluster number of 2. STEAP1/4 were co-clustered with CXCL1/8, IL-1 and IL-36 family proteins, as shown by the red boxes.
Figure E15. Clustering with gene cluster number of 3. STEAP1/4 were co-clustered with CXCL1/8, IL-1 and IL-36 family proteins, as shown by the red boxes.
Figure E16. Clustering with gene cluster number of 5. STEAP1/4 were co-clustered with CXCL1/8, IL-1 and IL-36 family proteins, as shown by the red boxes.
Figure E17. Clustering with sample cluster number of 2. The group of patient-only-samples (i.e. without healthy subjects) correlated with high expression of the STEAP1/4-CXCL1/8-IL1-IL36 group of genes, as shown by the circles.
Figure E18. Clustering with sample cluster number of 4. The group of patient-only-samples (i.e. without healthy subjects) correlated with high expression of the STEAP1/4-CXCL1/8-IL1-IL36 group of genes, as shown by the circles.
1