Fig 2: Effects of IVIg treatment in inflammatory infiltration in peripheral nerves. a Immunohistochemistry for CD68: After 3 and 8 weeks of BTZ treatment, both sciatic and caudal nerves show a massive macrophage infiltration which is almost completely abrogated by the preventive co-treatment with IVIg (BTZ + IVIg 3; BTZ + IVIg 8). In the therapeutic setting (BTZ + IVIg 4), a milder reduction of CD68+ infiltrating cells was achieved in both sciatic and caudal nerves. Scale bar 50 µm. b Representative immunohistochemistry for iNOS and ARG1 after 3 weeks of BTZ treatment. Most infiltrating macrophages (CD68+) are iNOS+ pro-inflammatory M1 type while a very limited amount of them are ARG1+ anti-inflammatory M2 type. Scale bar 50 µm

Fig 3: Astrocytes near MFB spheroids present the autophagy mechanisms needed to process the dopaminergic detritus. MFB astrocytes (a1) showed TH (a2) and LC3 (a3) immunoreactivity. a4 The LC3 and TH levels were highly correlated, and Pearson and Manders coefficients showed TH-LC3 colocalization. b1–b2 and b3–b4 show two examples of MFB astrocytes (cyan arrows; GFAP in cyan and DAPI in blue) near degenerating axons and spheroids of the MFB (green arrow in b1) and showing Lamp1 (yellow arrow in b2) and Lamp2 (yellow arrow in b4) immunoreactivity. DAergic cells (c1) of the substantia nigra with MitoDAT mitochondria (c2) expressed P62 (c3), a protein also found in spheroids (c4–c8). c4 A 3D example of a spheroid showing aggregation of mitochondria inside (c5). c6–c8 Consecutive slices of the aggregated mitochondria of c5 connected by P62 protein (white arrows). Cathepsin was found in the somata (black arrows in d1) and processes (white arrows in d2) of astrocytes of the degenerating MFB. d3–d8 Consecutive slices showing that the cathepsin immunoreactivity was particularly high in the astrocytic processes that penetrated or surrounded the mitochondrial aggregations of spheroids but not within the spheroids (white arrows). e–g Microglial cells (Iba1 in pink and DAPI in blue) located near spheroids or degenerating axons (black arrows) of the MFB neither accumulated TH (green) nor showed CD68 immunoreactivity. h TH-Iba1 cytofluorogram, and the Iba1-TH and Iba1-CD68 Pearson and Manders coefficients (n = 130) showing no statistical colocalization

Fig 4: Immunohistochemical staining for CD68-positive non-specific macrophages 2 weeks after implantation. Immunohistochemistry for CD68-positive cells. CD68 is one of the glycoproteins expressed in the monocytes and macrophages. Two weeks after implantation of MSC-CM, CD68-positive macrophages were not apparent in the MSC-CM group. Bar = 200 µm.

Fig 5: a-c Immunohistochemistry was performed on rat kidneys after syngeneic or allogeneic transplantation treated with CyA for 6 or 28 days, or CyA on alternating days until d28. Sections were analyzed using Histoquest® software after staining for rat CD3, CD20, and CD68. The mean number of CD3+/CD20+/CD68+cells/mm2 and standard error from at least 6 different animals per group is shown. Significance (Mann-Whitney U Test) is shown as * p < 0.05 compared to “SynTxd6”, and # p < 0.05 compared to “Txd28CyAalt”