Fig. 6: OXTR/ERK5 signaling maintains OXTR and CCL26 expression levels in OXTRHigh CAF. | Nature Communications

Fig. 6: OXTR/ERK5 signaling maintains OXTR and CCL26 expression levels in OXTRHigh CAF.

From: OXTRHigh stroma fibroblasts control the invasion pattern of oral squamous cell carcinoma via ERK5 signaling

Fig. 6

a saRNAs targeting the human OXTR gene promoter activate CAF markers and OXTR levels, and (b) graphical representation of OXTR promoter transcriptional activity. P = two-tailed t test. n = 3. c Results of ATAC–seq assay to assess chromatin accessibility of CAF activation and OXTR signaling-related genes in sa-NC and sa-OXTR groups of CAF1 for three independent experiments. d Representative IF staining and (e) quantification for four p-MAPK kinases in situ in Sa-NC or Sa-OXTR CAFs. Scale bar: 50 μm. P = multiple t test. n = 4. f Representative western blots of cytoplasmic and nuclear fractions of Sa-OXTR CAFs. n = 3 biologically independent experiments. g Graphical analyses of the promoter activity of OXTR/CCL26 of CAFs with sa-NC or sa-OXTR. P = two-tailed t test. n = 3. h Representative western blots of CAFs with sa-NC or sa-OXTR for endogenous ERK5 in in whole-cell lysates (WCL) or after immunoprecipitation with anti-ERK5 antibody for the proteins indicated. Four TFs in whole-cell lysates and IP were analyzed. i–l Mutations of potential binding sites of c-fos, c-myc on OXTR and CCL26 promoter and its transcription activity were analyzed by a dual-luciferase reporter assay system. P = multiple t test. n = 3. m Representative western blots and (n) quantification for the indicated proteins from CAFs lysates with or without sa-NC or sa-OXTR in WC) or immunoprecipitation with ERK5 for the CDC37/HSP90/ERK5 complex. P = multiple t test. n = 3. o Representative western blots and (p) quantification for the PKC/MEK5 signal complex for endogenous Gαq or β-arrestin in WCL or immunoprecipitation with Gαq or β-arrestin antibodies. n = 3. q Representative western blots and r quantification for knockdown of Gαq or β-arrestin by siRNA and determined the p-ERK5/OXTR/CCL26 levels. P = multiple t test. n = 3. s Representative (n = 3) IF images (scale bar: 50 μm) and (t) quantification of nuclear ERK5, MEK5 or CDC37 in sorted OXTRHigh or OXTRLow CAFs. P = two-tailed t test. u The OXTR signaling pathway diagram. Results are shown as mean and standard deviation (SD). All immunoblotting results are representative of three independent experiments. Source data are provided as a Source data file.

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