Fig. 7: Jumonji inhibition or KDM4A genetic knockdown alter the protein levels of key modulators of small cell lung cancer phenotypes in cells and in vivo.

A The SCLC marker Secretogranin-3 is decreased in KDM4A knockdown cell secreted media compared to Cas9 control H446 cells. A representative immunoblot is shown along with a section of total protein stain for the same gel. Quantification across two independent experiments normalized to total protein lysate is shown in the bar graph. B Western analysis validates the downregulation of INSM1 protein expression in H446 KDM4A KD cells, as well as in HCC4001, H510, and H2107 cells treated with Jumonji inhibitors, as indicated. GAPDH was used as a loading control. C NEUROD1 or ASCL1 Western blots show decreased protein in KDM4A KD or SD70 and JIB-04 treated cells. NEUROD1-high (H446, HCC4001) and ASCL1-high cells (H510, H2107) cells are shown. GAPDH as loading control. Note that in some cases the same gels were used to probe for INSM1 and NEUROD1 or ASCL1 thus sharing the same loading control. Quantifications are given in Fig. S7. D Western blots showing upregulation of H3K9me3 levels in H510 tumor nuclear extracts (n = 2 per treatment group) with RNA polymerase II used as a loading control. Quantification is shown on the right with lines connecting the DMSO control and SD70-treated mouse pairs. E Western blots showing decreased INSM1 and ASCL1 protein levels in H510 tumor lysates (n = 2 per treatment group) with GAPDH as a loading control. Quantifications are shown on the right with lines connecting DMSO control and SD70-treated mouse pairs. D, E dotted lines represent the place in the membrane where an extra control lane between DMSO and drug-treated samples was removed for simplicity of presentation.