Fig 1: Immunohistochemistry and Immunofluorescence of BCC. Immunohistochemical and immunofluorescent staining for GPR4 (GPR19), TDAG8 (GPR65), OGR1 (GPR68) and G2A (GPR132) in BCC tissue. (a–d) histochemical H&E staining, (e–h) immunohistochemical staining, inserted images present a 2 × larger field of view, (i–l) immunofluorescence staining, red: secondary antibody label, blue: DAPI. Scale bars correspond to 100 µm (a–l patient 20). Scores (bottom row) were assigned for ++: strong positive/positive reactions; +: weak positive/partial positive reaction; -: negative reaction. The expression of TDAG8 and G2A on the surface of BCC tumor cells is significantly increased, while the expression of GPR4 is weakly positive. No expression of OGR1 is detected. For additional stainings of other BCC, see Figures S13–S17.
Fig 2: IHC-Score results for different pH-GPCRs in common skin tumors. Fractional distribution of the scores (green bar, ++: strong positive/positive reaction; +, blue bar: weak positive/ partial positive reaction; -, red bar: negative reaction) for GPR4 (GPR19), TDAG (GPR65), OGR1 (GPR68) and G2A (GPR132) of immunohistochemically stained skin tumors (a) SCC, (b) BCC, (c) NCN, (d) MM. MM and NCN are further subclassified in epidermal and dermal portions. Numbers in bars indicate the occurrence of a respective score. The sum of all scores is 100%. Overall evaluation is indicated by a green, white or red box (beyond 100% scale): green box: overall positive reactions; red box: overall negative reactions; white box: mixed reactions without clear majority in favour of one specific reaction. Numbers in this box indicate the total number of samples investigated.
Fig 3: Immunohistochemistry and Immunofluorescence of NCN. Immunohistochemical and immunofluorescent staining for GPR4 (GPR19), TDAG8 (GPR65), OGR1 (GPR68) and G2A (GPR132) in NCN tissue. (a–d) histochemical H&E staining, (e–h) immunohistochemical staining, inserted images present a 2 × larger field of view, (i–l) immunofluorescence staining, red: secondary antibody label, blue: DAPI. Scale bars correspond to 100 µm (a–l patient 15). Scores (bottom row) were assigned for ++: strong positive/positive reactions; +: weak positive/partial positive reaction; -: negative reaction for the epidermal (left score) and the dermal (right score) portion. The NCN shows a partial positive expression of all GPCRs on the epidermal portions. There are even some epidermal tumor cells strongly expressed in the TDAG8. Regarding the dermal area GPR4, OGR1 and G2A are weak positive, while MM shows a significantly increased expression of TDAG8. For additional stainings of other NCN, see Figures S9–S12.
Fig 4: Immunohistochemistry and Immunofluorescence of SCC. Immunohistochemical and immunofluorescent staining for GPR4 (GPR19), TDAG8 (GPR65), OGR1 (GPR68) and G2A (GPR132) in SCC tissue. (a–d) histochemical H&E staining, (e–h) immunohistochemical staining, inserted images present a 2 × larger field of view, (i–l) immunofluorescence staining, red: secondary antibody label, blue: DAPI. Scale bars correspond to 100 µm (a–l: patient 1). Scores (bottom row) were assigned for ++: strong positive/positive reactions; +: weak positive/partial positive reaction; -: negative reaction. This SCC shows no expression of OGR1 and G2A in tumor cells, only several peritumoral lymphocytes appeared to be positive. The expression of GPR4 and TDAG8 is partial positive. For additional stainings of other SCC, see Figures S1–S4.
Fig 5: TMA-Score results for different pH-GPCRs in common skin tumors. Fractional distribution of the scores (green bar, ++: strong positive/positive reaction; +, blue bar: weak positive/ partial positive reaction; -, red bar: negative reaction) for GPR4 (GPR19), TDAG (GPR65), OGR1 (GPR68) and G2A (GPR132) of TMAs on different skin tumors (a) SCC, (b) BCC, (c) NCN, (d) MM. MM and NCN are further subclassified in epidermal and dermal portions. Numbers in bars indicate the occurrence of a respective score. The sum of all scores is 100%. Superscript numbers give information on scoring details: (1) strong positive either on the surface or in the deeper parts of the tumor tissue, (2) single tumor cells are strong positive, but the overall impression is weak positive, (3) large tumor cells appear strong positive, (4) partially strong positive and (5) weak positive. Overall evaluation is indicated by a green, white or red box (beyond 100% scale): green box: overall positive reactions; red box: overall negative reactions; white box: mixed reactions without clear majority in favour of one specific reaction. Numbers in this box indicate the total number of samples investigated. For additional information on the individual TMA score, see Supplementary Tables S2–S5.
Supplier Page from Abcam for Anti-GPCR G2A antibody