Fig. 3: Regulation of sGC subunit expression by Notch signalling in SCLC.

a Schematic of the working hypothesis. b Western blot for GUCY1B1 and synaptophysin (SYP) expression in SCLC cell lines (n = 3). c HEY1, GUCY1B1 and GUCY1A1 mRNA expression (RT-qPCR) of DMSO- or DAPT-treated H196 cells. n = 3, data are represented as mean ± SEM. P values from two-sided unpaired Student’s t-test. HEY1 p = 0.0001, GUCY1B1 p = 0.0002. d Western blot for GUCY1B1 and HES1 of DMSO- or DAPT-treated H196 cells. Quantification of GUCY1B1 (relative volume intensity, RVI) on the right, normalized to histone H3. n = 3, data are represented as mean ± SEM; p values from two-sided unpaired Student’s t-test. e Western blot for Notch1, HES1 and GUCY1B1 in sgNTA, sgNotch1-1 and sgNotch1-2 H1048 cells (n = 3). f Western blot for N1ICD, HES1 and GUCY1B1 in empty vector control and N1ICD-overexpressing H1048 cells. Quantification of GUCY1B1 (RVI) on the right, normalized to H3. n = 3, data are represented as mean ± SEM. P values from two-sided unpaired Student’s t-test. N1ICD p = 0.0008, HES1 p = 0.0009. g Western blot for GUCY1B1 and SYP in CDX17 and CDX17P NE and Non-NE ex vivo cultures (n = 2). h HEY1, GUCY1B1 and GUCY1A1 mRNA expression (RT-qPCR) of DMSO- or DAPT-treated CDX17P Non-NE cells. n = 3, data are represented as mean ± SEM. P values from two-sided unpaired Student’s t-test. i HEY1, GUCY1B1 and GUCY1A1 mRNA expression (RT-qPCR) of empty vector control or N1ICD-overexpressing CDX17P NE cells. n = 3, data are represented as mean ± SEM. P values from two-sided unpaired Student’s t-test. HEY1 p = 0.0005. j Notch1 ChIP-qPCR in H1048 cells overexpressing N1ICD. qPCR of RBPJ-binding sites in the GUCY1A1 and GUCY1B1 promoter. N1ICD binding to the HES1 promoter (positive control), binding to negative control region (negative control). n = 3, data are represented as mean ± SD. P values from two-sided unpaired Student’s t-test. See also Supplementary Fig. 3.