Skip to main content
. 2021 Jul 8;10:e64212. doi: 10.7554/eLife.64212

Figure 1. Mga inactivation in vivo leads to accelerated tumorigenesis.

(A) Schematic of lentiCRISPRv2cre vector and strategy used for in vivo Kras (K) and Kras, Tp53 (KP) experiments. (B) Kaplan–Meier curve for survival in sgMga (Cre-Cas9) vs Empty (Cre-Cas9 lacking sgMga) virus-treated Kras mice (p-value computed using Log-rank test). (C) Representative micrographs and (D) quantification of Ki67 immunostaining on lung tumor tissue from Kras-sgMga and Kras empty vector infected control mice (n = 5 mice per group. **p=0.0036. Calculated using Welch’s t-test assuming unequal variance). (E) Representative H and E staining of sgMga and Empty lungs from KP mice harvested 3 months post-intratracheal instillation. (F) Higher magnification of selected regions from (E) showing individual tumors in both groups. (G) Quantification of number of lesions per lobe and (H) number of lesions per grade in Empty (control) and sgMga vector treated animals (n = 5 lung lobes each from five mice per group. **p=0.001). (I) Representative images and (J) quantification of Ki67 staining to assess proliferation in KP Empty (control) and KP-sgMga lung tumors harvested 3 months post-infection (n = 32 tumors from four sgMga mice n = 21 tumors from two Empty (control) mice. ****p<0.0001. Calculated using Welch’s t-test assuming unequal variance). (K) Western blot and (L) immunofluorescent staining for MGA in cell lines derived from KP tumors. (M) Representative images and (N) quantification of Ki67 and MGA co-staining in an sgMga KP line upon ectopic expression of MGA (n = 23 FOV each from three independent experiments. ****p<0.0001). All p-values calculated using a two-sided Student’s t-test unless otherwise noted. Error bars represent SEM.

Figure 1.

Figure 1—figure supplement 1. Mga inactivation in vivo leads to accelerated tumorigenesis.

Figure 1—figure supplement 1.

(A) Oncoprint of TCGA and GENIE consortium data on genetic alterations in MGA in lung adenocarcinoma patients. (B) Kaplan–Meier curve for survival probability based on low and high expression of MGA in lung adenocarcinoma patients (KM plotter). (C) Synthego ICE analysis of representative tumor to show percentage of indels formed as a result of sgMga CRISPR treatment. Example shows a frameshift mutation (resulting from insertion of an additional Thymidine) at a frequency of 89.2%. (D) Percentage of tumors of each grade in Empty (control) vs. sgMga-infected Kras mice harvested at endpoint. (E) Representative H and E images of tumor grades observed in Kras mice. (F) Co-occurrence of MGA and TP53 alterations in TCGA and GENIE consortia data from lung adenocarcinoma patients. (G) Representative micrographs and (H) quantification of pan endothelial marker MECA32 in tumors from Empty vector and sgMga vector treated KP mice harvested 3 months post-intratracheal instillation (Empty: n = 9 tumors from three mice, sgMga: n = 21 tumors from two mice. ***p=0.0003 using Welch’s t-test assuming unequal variance). (I) Quantification of MGA intensity in Empty (sg-control), sgMga1 and sgMga3 KP tumor cell lines (n = 5 FOV for all groups). Error bars represent SEM.