MOESM4 of Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction

Additional file 4: Table S4. List of primers used for qRT-PCR. This table lists the primers used for the qRT-PCR studies

MOESM3 of Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction

Additional file 3: Table S3. List of antibodies used for ICC and flow cytometry. This table provides a list of antibodies used for the ICC and flow cytometry studies

MOESM2 of Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction

Additional file 2: Table S2. List of BD Biosciences Flex Sets used for CBA. This table lists the BD Biosciences Flex Sets used for the CBA studies

MOESM1 of Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction

Additional file 1: Table S1. In vitro culture conditions of brain pericytes in selected publications. This table lists previous publications describing in vitro pericyte cell culture conditions

Additional file 6: Figure S6. of Interferon-γ blocks signalling through PDGFRβ in human brain pericytes

Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh), TNFα (5 ng/mL), or IL-1β (1 ng/mL). After 48 h of cytokine treatment, cells were treated with either vehicle or PDGF-BB (10 ng/mL) to measure the PDGF-BB-induced proliferative response (A, B). This was done in two ways: after 96 h total treatment, cells were fixed, labelled with a Ki67 antibody and Hoechst (A, C); alternatively, EdU was added to measure cell proliferation over the final 24 h of the experiment (B, C). Positive cells of the total cells measured by Hoechst were quantified and plotted as mean ± s.e.m. (n = 3), ****(p < 0.0001), ***(p < 0.001), *(p < 0.05) from a two-way ANOVA. (TIF 1034 kb

Additional file 4: Figure S4. of Interferon-γ blocks signalling through PDGFRβ in human brain pericytes

Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh), TNFα (5 ng/mL), or IL-1β (1 ng/mL). After 96 h total treatment, cells were serum starved for 2 h and then treated with vehicle (−) or PDGF-BB (100 ng/mL) for 30 min. PDGFRβ puncta were quantified using MetaXpress™ software and normalized to cell number and vehicle control and plotted as mean ± s.e.m. (n = 3), ***(p < 0.001), *(p < 0.05) (two-way ANOVA). Note: Control data are from the same experiments as Fig. 3g. (TIF 723 kb

Additional file 3: Figure S3. of Interferon-γ blocks signalling through PDGFRβ in human brain pericytes

Repeats of additional cases from Fig. 3(b–e): Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh) or IFNγ (1 ng/mL). After 96 h total treatment, cells were serum starved for 2 h and then treated with vehicle (−) or PDGF-BB (100 ng/mL) for 30 min as in Fig. 3. (a, c) Representative western blots of treated pericyte from two additional cases. (B, D) Bands were quantified with Image Studio™ and normalized to vehicle control. p-PDGFRβ was normalized to total PDGFRβ, and p-Akt and p-ERK were normalized to total Akt and ERK, respectively. (TIF 2398 kb

Additional file 2: of Unique and shared inflammatory profiles of human brain endothelia and pericytes

Full secretome profiler dataset from Fig. 5. Pericytes or endothelia from two different cases were treated with either vehicle, or IL-1β (10 ng/mL) for 24 h before media were harvested and secretions analysed as per Fig. 5. Values represent intensity measurements from each secretion spot (two spots per secretion) from both biological replicates, from all treatment groups (Vehicle/Pericyte, IL-1β/Pericyte, Vehicle/Endothelial, IL-1β/Endothelial). Data are given in an Excel spreadsheet. (XLSX 41 kb

Additional file 7: Figure S7. of Interferon-γ blocks signalling through PDGFRβ in human brain pericytes

(A, B) Pericytes were treated for four consecutive days (once every 24 h) with either vehicle (Veh), TNFα (5 ng/mL), or IL-1β (1 ng/mL). After 48 h of cytokine treatment, cells were treated with either vehicle or PDGF-BB (10 ng/mL) to measure PDGFRβ and αSMA expression by immunocytochemistry. Quantification of PDGFRβ (A) and αSMA (B) staining, mean ± s.e.m. (n = 3), ****(p < 0.0001), ***(p < 0.001), *(p < 0.05) (two-way ANOVA). (C) Pericytes were treated for three or four consecutive days (once every 24 h) with either vehicle (Veh), TNFα (5 ng/mL), or IL-1β (1 ng/mL). After 48 h, cells were treated with PDGF-BB (10 ng/mL) for either 24 or 48 h. Western blot band intensity of PDGFRβ, αSMA, and GAPDH were quantified, normalized to GAPDH, and plotted as mean ± s.e.m. (n = 3), and differences were not significant (two-way ANOVA). (TIF 1163 kb

Additional file 1: Table S1. of TGF-beta1 regulates human brain pericyte inflammatory processes involved in neurovasculature function

List of antibodies used for immunocytochemistry. List of antibodies, suppliers and dilutions used for immunocytochemistry studies. (16.9 kb