Figure 3.
VA-lip HSP47 ameliorate fibrosis of the skin and salivary glands in chronic GVHD. (A-F) Mice were transplanted as in Figure 1. A group of allogeneic recipients were treated with VA-lip HSP47 or VA-lip containing scramble siRNA at 3 times per week from day +2 to day +41, and skin samples were harvested on day +42 after BMT. (A) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (B) Numbers of positive pixels for HSP47 in syngeneic (n = 9), allogeneic controls (n = 13), and allogeneic recipients treated with VA-lip containing scramble siRNA (n = 12) or VA-lip HSP47 (n = 13) from 2 of 3 independent experiments were combined and shown as means ± SEM. (C) Immunofluorescent images for RBP1 (green) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (D) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. Skin thickness (E) and amount of collagen deposit (F) in syngeneic (n = 19), allogeneic controls (n = 28), and allogeneic recipients treated with VA-lip containing scramble siRNA (n = 15) or VA-lip HSP47 (n = 21) from 5 independent experiments were combined and shown as means ± SEM. (G-H) Submandibular glands were harvested on day +42 after BMT. (G) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. The area in the white rectangle is magnified and is shown on the right side of the original image. (H) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. (I-M) Lethally irradiated B6 mice were transplanted with G-CSF–mobilized splenocytes from B6 (Syn, n = 10) or BALB/c (Allo, n = 21) donors. Allogeneic recipients were treated with VA-lip HSP47 (n = 7) or diluent (n = 14) at 3 times per week from day +2 to day +41, and skin samples were harvested on day +42 after BMT. (I) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (J) Numbers of positive pixels for HSP47 in syngeneic, allogeneic controls, and allogeneic recipients treated with VA-lip HSP47. (K) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. Skin thickness (L) and amount of collagen deposit (M) in syngeneic, allogeneic controls, and allogeneic recipients treated with VA-lip HSP47. Data from 2 independent experiments were combined and are shown as means ± SEM. *P < .05; **P < .01.

VA-lip HSP47 ameliorate fibrosis of the skin and salivary glands in chronic GVHD. (A-F) Mice were transplanted as in Figure 1. A group of allogeneic recipients were treated with VA-lip HSP47 or VA-lip containing scramble siRNA at 3 times per week from day +2 to day +41, and skin samples were harvested on day +42 after BMT. (A) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (B) Numbers of positive pixels for HSP47 in syngeneic (n = 9), allogeneic controls (n = 13), and allogeneic recipients treated with VA-lip containing scramble siRNA (n = 12) or VA-lip HSP47 (n = 13) from 2 of 3 independent experiments were combined and shown as means ± SEM. (C) Immunofluorescent images for RBP1 (green) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (D) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. Skin thickness (E) and amount of collagen deposit (F) in syngeneic (n = 19), allogeneic controls (n = 28), and allogeneic recipients treated with VA-lip containing scramble siRNA (n = 15) or VA-lip HSP47 (n = 21) from 5 independent experiments were combined and shown as means ± SEM. (G-H) Submandibular glands were harvested on day +42 after BMT. (G) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. The area in the white rectangle is magnified and is shown on the right side of the original image. (H) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. (I-M) Lethally irradiated B6 mice were transplanted with G-CSF–mobilized splenocytes from B6 (Syn, n = 10) or BALB/c (Allo, n = 21) donors. Allogeneic recipients were treated with VA-lip HSP47 (n = 7) or diluent (n = 14) at 3 times per week from day +2 to day +41, and skin samples were harvested on day +42 after BMT. (I) Immunofluorescent images for HSP47 (red) with DAPI (blue) counterstaining. Magnification, ×40. Scale bar, 50 µm. (J) Numbers of positive pixels for HSP47 in syngeneic, allogeneic controls, and allogeneic recipients treated with VA-lip HSP47. (K) Representative images of MT staining. Magnification, ×20. Scale bar, 50 µm. Skin thickness (L) and amount of collagen deposit (M) in syngeneic, allogeneic controls, and allogeneic recipients treated with VA-lip HSP47. Data from 2 independent experiments were combined and are shown as means ± SEM. *P < .05; **P < .01.

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