Additional file 2: Figure S2. of Evidence for an early innate immune response in the motor cortex of ALS

MCP1+ cells and CCR2+ cells are present in the motor cortex and they are in close proximity to CSMN in the MCP1-CCR2-hSOD1G93A mice. (a) Representative image showing very few MCP1+ cells in the motor cortex of MCP1-CCR2-WT mice. (b) Representative image showing increased numbers of MCP1+ cells in the motor cortex of MCP1-CCR2-hSOD1G93A mice. MCP1+ cells express microglia marker CD11b. Insets enlarged to the right (b’-b”’). (c) Experimental design depicting retrograde transduction of CSMN approach using AAV-eGFP in the MCP1-CCR2-WT and MCP1-CCR2-hSOD1G93A mice. AAV2-eGFP was injected into the CST of mice at P30, and tissue was collected at P60. (d, e) Representative images show MCP1+ cells near transduced CSMN (eGFP+) in the layer V of the motor cortex (d, e) and in the layer II/III of the motor cortex (f, g) in MCP1-CCR2-hSOD1G93A mice. (h, j) Representative images show MCP1+ cells expressing phagocytic marker CD68 and their interaction with transduced CSMN in the layer V of motor cortex in the MCP1-CCR2-hSOD1G93A mice. (k-n) Representative image showing CCR2+ cells in layer II/III of motor cortex co-localizing with monocyte marker CD45 and infiltrating monocyte marker Ly6C. Scale bar:s: a,b,d-g =20 μm; k-n = 10 μm. (PDF 1521 kb

Additional file 3: Figure S3. of Evidence for an early innate immune response in the motor cortex of ALS

MCP1+ cells express neither Arginase 1 (Arg1) nor inducible nitric oxide synthase (iNOS) in the MCP1-CCR2-hSOD1G93A mice. (a) Representative images of Arg1+ cells (arrowheads) and MCP1+ cells (arrows) in the liver of MCP1-CCR2- hSOD1G93A mice 6 h post LPS I.P. injection (positive control). (b) Representative images of 2° only for Arg1 (negative control) and MCP1+ cells (arrows) in the liver of MCP1-CCR2- hSOD1G93A mice 6 h post LPS I.P. injection. (c) Representative images of MCP1+ cells (arrows) in the spleen of MCP1-CCR2- hSOD1G93A mice 6 h post LPS I.P. injection (positive control) show co-localization with iNOS (arrows). (d) Representative images of 2° only for iNOS (negative control) and MCP1+ cells (arrows) in the spleen of MCP1-CCR2- hSOD1G93A mice 6 h post LPS I.P. injection. (e) Experimental design depicting retrograde transduction of CSMN approach using AAV-eGFP in the MCP1-CCR2-WT and MCP1-CCR2-hSOD1G93A mice. AAV2-eGFP was injected into the CST of mice at P30, and tissue was collected at P60. (f-g) Representative images of the layer II/III of motor cortex show lack of co-localization of MCP1+ cells with Arg1 in MCP1-CCR2-WT mice (f) and MCP1-CCR2- hSOD1G93A mice (g). (h-i) Representative images of the layer II/III of motor cortex show lack of co-localization of MCP1+ cells with iNOS in MCP1-CCR2-WT mice (h) and MCP1-CCR2- hSOD1G93A mice (i). Scale bar = 10 μm. (PDF 961 kb