Fig. 4

MANF protein expression is induced in Iba1 + cells of the Nestin^Cre/+::Manf^fl/fl knockout mouse brain after ischemic stroke. Anti-MANF immunostaining of Manf^fl/fl control (a, c, e) and Nestin^Cre/+::Manf^fl/fl knockout (b, d, f) mouse brain at 14d after permanent dMCAo. g-h Anti-rabbit IgG immunostaining of the same brain as in a, b to show the specificity of the rabbit anti-MANF antibody. i Average infarct area at 14d after permanent dMCAo in Manf^fl/fl wild type and Nestin^Cre/+::Manf^fl/fl knockout mouse. a, b, g, h: scale bar is 500 µm. c-f: scale bar is 100 µm. j-m Anti-MANF and anti-Iba1 double immunofluorescence staining of the peri-infarct cortex of Manf^fl/fl control (j, k) and Nestin^Cre/+::Manf^fl/fl knockout (l, m) mice at 14d after permanent dMCAo. Scale bar is 10 µm. n Endogenous MANF serum levels are not altered after permanent cerebral ischemia. Endogenous MANF levels were measured from wild type mouse serum with mouse MANF ELISA at different time points after permanent dMCAo, n = 3–6 per group. The values are expressed as mean ± SD. dMCAo = distal middle cerebral artery occlusion