Fig 2: Investigation of mambalgin-2 interaction with the ASIC1a (a,b), a-ENaC (c,d), ?-ENaC (e,f), and ASIC3 (g,h) subunits in mel P cells: localization of fluorescently-labelled mambalgin-2/CF647 (red) and Alexa488-labeled ASIC1 (a), a-ENaC (c), ?-ENaC (e), and ASIC3 (g) subunits (green). Nuclei were stained by Hoechst 33342 (blue), scale bar 10 µm. Western blot analysis of the ASIC1 (b), a-ENaC (d), ?-ENaC (f), and ASIC3 (h) subunits extraction from membrane fraction of mel P cells by affinity chromatography on NHS-sepharose resin coupled with mambalgin-2 (n = 3).

Fig 3: Bioinformatic analysis of the ACCN2, ACCN1, ACCN3, ACCN4, SCNN1A, and SCNN1G expression in healthy people and patients with lung adenocarcinoma from the GTEX and TCGA LUAD databases. (A) Analysis of the gene expression in healthy lung biopsies (GTEX study) and in samples of lung adenocarcinoma surrounding tissue, and lung adenocarcinoma specimen (TCGA LUAD study). Data are presented as normalized gene expression ± SEM; *** (p < 0.001) indicates significant difference between the data groups by one-way ANOVA followed by Tukey’s post-hoc test. (B, C) Kaplan–Meier analysis of the correlation between survival of the patients with lung adenocarcinoma at I (B) and II–IV stages (C) and different expressions of the ACCN2, ACCN1, ACCN3, ACCN4, SCNN1A, and SCNN1G genes. * (p <0.05) indicates significant difference between the overall survival prognosis for patients with the high (above median) and low (below median) gene expressions according to the log-rank test, n.s. - no significant difference (p>0.05).

Fig 4: Bioinformatic analysis of the ACCN2, ACCN1, ACCN3, ACCN4, SCNN1A, and SCNN1G expression in healthy people and patients with non-glabrous primary and metastatic melanoma from the GTEX and TCGA SKSM databases: (a) analysis of the gene expression in healthy skin biopsies (GTEX study) and in samples of primary and metastatic melanoma (TCGA SKSM study). Data are presented as normalized gene expression ± SEM, *** (p < 0.001) indicates significant difference between the data groups by One-Way ANOVA followed by Dunnets’ hoc test; (b,c) Kaplan–Meier analysis of survival of the patients with primary (b) and metastatic (c) melanoma and different expression of the ACCN2 gene. * (p < 0.05) indicates significant difference between the survival prognosis for patients with the high (above median) and low (below median) ACCN2 expression according to log-rank test.

Fig 5: qPCR analysis of the ACCN2, ACCN1, ACCN3, ACCN4, SCNN1A, and SCNN1G expression in A549, WI-38, and HLF cells. Gene expression was normalized to the ß-ACTIN, GPDH, and RPL13a housekeeping genes and presented as relative mRNA level ± SEM (n = 6). * (p < 0.05), ** (p < 0.01), and *** (p < 0.001) indicate significant difference between the data groups according to one-way ANOVA followed Tukey’s post-hoc test, n.s. - no significant difference (p>0.05).