Fig 2: Shikonin promotes leukemia cell differentiation. HL-60 cells were treated with shikonin for 48 h. (A) Cell viability was detected by MTT assay. (B) Cell morphology was detected by Wright Giemsa staining. (C) The cell volume distribution was detected by the Holomonitor system. (D) The expression of CD34 and CD11b was detected by immunofluorescence in HL-60 cells. (E) CD34 and CD11b protein expression was detected by Western blot. The data were presented as the mean ± SD (n = 3). * p < 0.05; ** p < 0.01 compared to the untreated control.

Fig 3: Expression of renalase in glomerular macrophages in active proliferative LN.(A) Left panel—kidney immunohistochemical staining for renalase of a control subject (20X); Right panel kidney immunohistochemical staining for renalase of an active LN patient (20X), arrows indicate representative renalase expression in glomeruli; (B) immunofluorescent staining of representative kidney sections of a patient with active LN, Left panel–staining with anti-CD68 antibody, Middle left- staining with anti-renalase antibody, Middle right -: nuclear staining, Right panel- merged picture; (C) immunofluorescent staining of representative kidney sections of a patient with active LN, Left panel—staining with anti-CD34 antibody, Middle left—staining with anti-renalase antibody, Middle right-: nuclear staining, Right panel—merged picture; (D) immunofluorescent staining of THP–1 cells in culture; Left panel—staining with anti-CD68 antibody, Middle left—staining with anti-renalase antibody, Middle right-: nuclear staining, Right panel—merged picture.

Fig 4: Manifestations of hypoxia, angiogenesis, and oxidative stress in the rat prostate. (a) Representative IHC photos of CD34 of the rat prostate samples (upper, magnification ×100; lower, magnification ×200). (b) The counting result of MVD. (c) Representative western blot results of HIF-1a, VEGF, and eNOS protein expressions. ß-Actin served as an internal control. (d) The relative optical density quantification of the proteins mentioned above according to western blot. (e) Relative mRNA expression levels of HIF-1a, VEGF, and eNOS to ß-actin by QRT-PCR. (f) Representative fluorescence photos of ROS in situ of the prostate samples of the rats (upper, red, simple ROS staining, magnification ×200; lower, merge of ROS and DAPI, magnification ×200). (g) The fluorescence intensity of ROS. (h) SOD activity of the rat prostates. (i) MDA content of the rat prostates. Each bar in (b), (d), (e), (g), (h), and (i) represents the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Fig 5: Overexpression of HMGB3 correlates positively with MVD in NPC xenografts.A, H Images of corresponding tumour xenografts. B, I Tumour volume in the two groups. C, J The weights of the excised xenografts. D, K Representative images of IHC for CD34 and HMGB3 in tissues collected from two groups of NPC xenografts. E, L Quantification of IHC staining score for HMGB3 expression. F, M Statistics of microvessel density. G, N Spearman correlation between HMGB3 score and MVD in tumour xenografts. Pearson correlation coefficient (r2) and P value are shown. The scale bar in 400× images represents 50 µm. The red arrows indicate microvessels. Mean ± SD, **P < 0.01, ***P < 0.001, ****P < 0.0001, student’s test.