Rabbit anti-BCL-2 Polyclonal Antibody

Design of therapy protocol. Group A received saline. Group B received 18.5 MBq of 177Lu-EB-RGD targeted radionuclide therapy (TRT) by intravenous injection. Group C received anti-PD-L1 mAbs (αPD-L1 mAb) immunotherapy at a dose of 10mg/kg by intraperitoneal injection (d 0, 3 and 6). Group D received concurrent combined therapy with 177Lu-EB-RGD (d 0) and αPD-L1 mAb (d 1, 4 and 7), and group E received sequential combined therapy with 177Lu-EB-RGD (d 0) and αPD-L1 mAb (d 11, 14 and 17).

(A) Projection SPECT images of MC38 tumor bearing mice at 4, 24, 48, 72 and 96 h after injection of 18.5 MBq 177Lu-EB-RGD (n = 4). (B) Projection SPECT images of MC38 tumor bearing mice at 1 and 4 h after injection of 18.5 MBq 177Lu-RGD (n = 4). (C) Area under the curves (AUC) of 177Lu-EB-RGD (blue) and 177Lu-RGD (purple) calculated from the radioactivity accumulation in the tumor over time. Arrows indicate tumor location.

(A) Design of therapy protocol. (B) Body weight change. (C) Tumor volume. (D) Survival of mice (9-10/group) treated with saline (group A), 177Lu-EB-RGD (group B), αPD-L1 mAb (group C), concurrent combined therapy with 177Lu-EB-RGD and αPD-L1 mAb (group D) and sequential combined therapy with 177Lu-EB-RGD and αPD-L1 mAb (group E) at different days after treatment. (E) Projection PET images of MC38 tumor bearing mice injected with 18F-FDG 3 d after treatment with saline (group A), 177Lu-EB-RGD (group B), αPD-L1 mAb (group C) and combined therapy with 177Lu-EB-RGD and αPD-L1 mAb (group D). White arrows indicate the tumor location. (F) PET quantification of the images in Fig 4E. *P < 0.05, ** P < 0.01

Immunofluorescence staining of excised tumors for CD31 (vascularity), Ki-67 (proliferation), TUNEL (apoptosis), and hematoxylin and eosin (H&E) after treatment with saline (group A), 177Lu-EB-RGD (group B), αPD-L1 mAb (group C), concurrent combined therapy with 177Lu-EB-RGD and αPD-L1 mAb (group D) and sequential combined therapy with 177Lu-EB-RGD and αPD-L1 mAb (group E). Tumor samples from groups A-D were collected at d 7 after treatment, and samples from group E were collected at d 14 after treatment (αPD-L1 mAb starting on d 11 of TRT).

(A) Multiplexed immunofluorescence staining for PD-L1 (pink), CD11b+ (red), CD45+ (green), CD8+ (yellow) and DAPI (blue) in MC38 tumors, which were measured 4 d (left), 7 d (middle) and 14 d (right) after treatment with saline (control), 177Lu-EB-RGD targeted radionuclide therapy (TRT), concurrent combined therapy with 177Lu TRT and αPD-L1 mAb (day 4 and 7) or sequential combined therapy with 177Lu TRT and αPD-L1 mAb (day 14). Scale bar is 200μm (magnification, × 20). (B-D) Quantified results for multiplexed immunofluorescence staining presented in Figure 7A at different time points. *P < 0.05, **P < 0.01, ***P<0.001

(M) Western blot images and Bax/Bcl-2 ratios (n = 3).