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1.
Figure 2

Figure 2. Alginate bead release kinetics of rLOX-PP.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Western blot of known amounts of rLOX-PP ranging from 0.1 to 10 ng of rLOX-PP; (B) Western blot of rLOX-PP-bead supernatants collected at different intervals; (C) calculated cumulative release of rLOX-PP; (n = 4; *, p<0.001).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
2.
Figure 8

Figure 8. rLOX-PP persists in treated tumors.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

Immunostaining of tumors after sacrifice with LOX-PP antibody or non-immune IgG control from mice after (A) direct i.t. injection of rLOX-PP compared to PBS or (B) after implantation of empty or rLOX-PP-containing alginate beads. Scale bars = 0.005 mm. Arrows mark some positive-stained cells.

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
3.
Figure 9

Figure 9. LOX-PP inhibits Ras effector Erk1/2 phosphorylation in vivo.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

Western blot analysis of 42 and 44 kDa phospho-Erk, total Erk1/2 and 36 kDa beta-actin from tumor extracts after (A) direct i.t. injections of rLOX-PP (L) compared to PBS (P); (B) after implantation of empty (EB) and rLOX-PP (LB) alginate beads; (C) the corresponding quantifications are shown (*, p<0.05; n = 3).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
4.
Figure 4

Figure 4. Proliferation marker Ki-67 is decreased by rLOX-PP treatment.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Immunostaining with Ki-67 antibody or non-immune IgG control tumors after (A) direct i.t. injection of rLOX-PP compared to PBS and (B) after implantation of empty and rLOX-PP alginate beads. Black arrows mark some positive-stained cells; the white arrows point to an inset containing an enlarged image of a stained cell. (C) The corresponding quantifications are shown (***, p<0.001; n = 3). Scale bar = 0.005 mm.

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
5.
Figure 7

Figure 7. Apoptosis marker TUNEL is increased by rLOX-PP treatment.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) TUNEL staining of tumors treated i.t. with rLOX-PP, or tumors injected with negative control PBS; (B) tumors treated with surgically implanted empty alginate beads or rLOX-PP alginate beads. Black arrows mark some positive-stained cells; the white arrows point to an inset containing an enlarged image of a stained cell. Scale bar = 0.005 mm for 400× and 0.002 mm for 1000×; (C) the corresponding quantifications (***, p<0.001; n = 3). Arrows mark some positive-stained cells.

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
6.
Figure 6

Figure 6. Apoptosis marker active caspase 3 is increased by rLOX-PP treatment.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Immunostaining of tumors with active caspase 3 antibody, or non-immune IgG control. (A) i.t. rLOX-PP- or PBS injected tumors, and (B) tumors treated with surgically implanted empty alginate beads or rLOX-PP/alginate beads. Black arrows mark some positive-stained cells; the white arrows point to an inset containing an enlarged image of a stained cell. Scale bar = 0.005 mm for 400× and 0.002 mm for 1000×; (C) the corresponding quantifications (***, p<0.001; n = 3).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
7.
Figure 3

Figure 3. rLOX-PP incorporated in alginate beads inhibits NF639 xenograft growth in mice compared to empty beads.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Subcutaneous 100 mm3 xenografts of NF639 breast cancer cells were established in mice. rLOX-PP/alginate or empty beads were implanted on day 15 (arrow). Tumors were monitored by caliper measurements (n = 10; *, p<0.05, ***, p<0.01). (B) The tumor growth rates were determined from measurements made beginning on day 18 and were derived from linear regression analyses of the data; (C) tumor weights determined at sacrifice on day 25 (n = 10; *, p<0.05).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
8.
Figure 1

Figure 1. Injection of rLOX-PP inhibits NF639 xenograft growth in mice.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Subcutaneous xenografts of NF639 breast cancer cells were injected once daily with 10 micrograms rLOX-PP for 5 days beginning on day 17 (arrows). Tumor volumes were monitored by caliper measurements (n = 10; *, p<0.05); (B) The tumor growth rates were determined from measurements made beginning on day 18 and were derived from linear regression analyses of the data. (C) Tumor weights at sacrifice on day 25 (p = 0.4, n = 10).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.
9.
Figure 5

Figure 5. Proliferation marker phosphorylated histone H3 is decreased by rLOX-PP treatment.. From: Recombinant Lysyl Oxidase Propeptide Protein Inhibits Growth and Promotes Apoptosis of Pre-Existing Murine Breast Cancer Xenografts.

(A) Immunostaining with phospho- histone H3 antibody or nonimmune IgG control tumors after (A) direct i.t. injection of rLOX-PP (L) compared to PBS (P) and (B) after implantation of empty (EB) and rLOX-PP (LB) alginate beads. Black arrows mark some positive-stained cells; the white arrows point to an inset containing an enlarged image of a stained cell. Scale bars correspond to 0.005 mm for 400× and 0.002 mm for 1000×. Arrows mark some positively stained cells. (C) Western blot analyses of 17 kDa phospho- histone H3, 36 kDa beta-actin and the corresponding quantifications of protein extracts from tumors after (C) direct i.t. injections of rLOX-PP compared to PBS and (D) after implantation of empty and rLOX-PP alginate beads (*, p<0.01; n = 3).

Manish V. Bais, et al. PLoS One. 2012;7(2):e31188.

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