Fig. 6: P4 cells exhibit dysregulated Mitf genes and upregulated Gpnmb promoter. | Communications Medicine

Fig. 6: P4 cells exhibit dysregulated Mitf genes and upregulated Gpnmb promoter.

From: Acquired resistance to immune checkpoint therapy is caused by glycoprotein non-metastatic melanoma protein B signal cascade

Fig. 6

qRT-PCR analysis of Mit family (Mitf, Tfe3, Tfeb, and Tfec) (a) and Sox10 (b) in P0 vs. P4 cells (mean ± SEM, n = 3). c Proportion of Mitf isoform transcripts in P0 vs. P4 cells is calculated and expressed in Pie chart. d RT-PCR analysis of Mitf isoforms in varying cell lines (P0, P4, B16 melanoma, and Raw macrophages), run on 1.5% agarose gel/ethidium bromide. The bp size of PCR products is shown at the right of the figure. e P0 or P4 cells were transfected with increasing doses of pGL3b (basic promoter-Luc), pG-Gpnmb-p (Gpnmb promoter-linked pGL3b), pG-Gpnmb-p-ΔA (MITF-A site-deleted Gpnmb promoter), pG-Gpnmb-p-ΔΒ (MITF-B-deleted), or pG-Gpnmb-p-ΔAB (both A and B sites-deleted). Luciferase (Luc) activities were measured and expressed as Luc/Renilla × 100 (n = 3, average ± SD). Representative of three experiments. f Heatmap analyses of expression of 43 Mitf-target genes in P0 vs. P4 cells (three different batches per sample) are shown using RNA-seq data (Supplementary Data 2 and 3). p values in (a, b) are shown using Student’s t-test. Adjusted p value is indicated in parenthesis, using the Benjamini–Hochberg procedure.

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