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. 2022 Sep 16;82(22):4261–4273. doi: 10.1158/0008-5472.CAN-21-3214

Figure 5.

Figure 5. Rbms3 loss cooperates with EGFRL858R to accelerate malignant lung adenocarcinoma. A–C, Images of harvested mouse lung sections following necropsy analyses stained with hematoxylin and eosin (H&E) 11 weeks after initiation with 1×105 pfu lenti-CRE, followed by continuous administration of doxycycline chow to induce EGFRL858R expression. CRISPR/CAS9-mediated genome editing was used in B and C to edit Rbms3 in vivo. Genotype of each experimental group was: A, sgRbms3-CRE virus in SPC::CRE-ERT2/+; Rosa26CAGs-LSL-rTTa3; EGFRL858R (SRE) mice. B, sgNT-CRE virus in or SPC::CRE-ERT2/+; Rosa26CAGs-LSL-rTTa3; EGFRL858R; H11LSL-CAS9/+ (SREC) mice. C, sgRbms3-CRE virus in SREC mice. Black bar in bottom left of each panel represents a 1,000 μm, scale bar. D, Quantification of tumor burden from genotypes in B compared with C. N = 5 mice per group. The mean is graphed, and error bars represent SEM. Statistical analysis was conducted using a paired t test; ****, P < 0.0001. E, Representative images of IHC on FFPE lung tissue sections from SREC mice initiated with either sgNT- or sgRbms3-CRE and stained with pEGFR (pY1068) or pERK (pT202; Y204) shown at ×20 magnification. Scale bar shown in black at the bottom left corner of each image represents 50 μm.

Rbms3 loss cooperates with EGFRL858R to accelerate malignant lung adenocarcinoma. A–C, Images of harvested mouse lung sections following necropsy analyses stained with H&E 11 weeks after initiation with 1×105 pfu lenti-CRE, followed by continuous administration of doxycycline chow to induce EGFRL858R expression. CRISPR/CAS9-mediated genome editing was used in B and C to edit Rbms3 in vivo. Genotype of each experimental group was: A, sgRbms3-CRE virus in SPC::CRE-ERT2/+; Rosa26CAGs-LSL-rTTa3; EGFRL858R (SRE) mice. B, sgNT-CRE virus in or SPC::CRE-ERT2/+; Rosa26CAGs-LSL-rTTa3; EGFRL858R; H11LSL-CAS9/+ (SREC) mice. C, sgRbms3-CRE virus in SREC mice. Scale bar, 1,000 μm. D, Quantification of tumor burden from genotypes in B compared with C. N = 5 mice per group. The mean is graphed. Error bars, SEM. Statistical analysis was conducted using a paired t test; ****, P < 0.0001. E–H, Representative images of IHC on FFPE lung tissue sections from SREC mice initiated with either sgNT- or sgRbms3-CRE and stained with pEGFR (pY1068) or pERK (pT202; Y204) shown at ×20 magnification. Scale bar, 50 μm.