Skip to main content
. Author manuscript; available in PMC: 2022 Aug 3.
Published in final edited form as: Nat Cancer. 2022 Feb 3;3(2):188–202. doi: 10.1038/s43018-021-00316-3

Fig. 4 |. SETD2 loss-of-function induces MMP1 expression to promote ccRCC metastasis.

Fig. 4 |

a, The MMP1 mRNA was assessed in the indicated JHRCC12 cells by qRT-PCR. Data were normalized against β-Actin (mean ± s.d., n = 3 independent experiments). b, ATAC-seq tracks at the MMP1 locus in the indicated JHRCC12 cells. c, The indicated JHRCC12 cells were assessed by ChIP-qPCR for the promoter and intron 7 of MMP1. Data shown are the percent input (mean ± s.d., n = 3 independent experiments). d, CAKI-2 cells were transiently transfected with either pGL2-pro or pGL2-pro containing the DNA fragment from the ATAC-seq peak at the intron 7 of MMP1 together with pRL-SV40. The firefly and Renilla luciferase activities were assessed and normalized (mean ± s.d., n = 3 independent experiments). e, The MMP1 mRNA was assessed in JHRCC12 cells transfected with dCas9–KRAB–MeCP2 and the indicated sgRNAs as in a. f, Whole cell lysates (WCL) and histone fractions from the indicated JHRCC12 cells were assessed by immunoblots. g, The MMP1 mRNA was assessed in the indicated JHRCC12 cells by qRT-PCR as in a. h, The indicated JHRCC12 cells were assessed by immunoblots. The asterisk denotes a cross-reactive band. Cellular proliferation was assessed by CellTiter-Glo assays. i, The indicated luciferase-transduced JHRCC12 cells were injected into subrenal capsules of unilateral kidneys of NSG mice to establish orthotopic xenografts. The weight of kidney tumors in each mouse was estimated by subtracting the weight of kidney without orthotopic implantation from that of kidney received subcapsular injection of the indicated JHRCC12 cells after 5–6 weeks. Data shown are mean ± s.d. (n = 4 mice for sgLacZ and n = 8 mice for sgMMP1). j, Bioluminescence images of athymic nude mice at the indicated times after intracardiac injection of the indicated luciferase-transduced JHRCC12 cells. k, Quantification of bioluminescence shown in j (mean ± s.d., n = 5 mice for each group). P = 0.0055 (two-way ANOVA). P values in a, c, d, e and g were determined by two-tailed unpaired Student’s t-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.